Tuesday, February 28, 2017

Stem Cells

What Are Stem Cells?

Stem cells are unique cells of the body in that they are unspecialized and have the ability to develop into several different types of cells. They are different from specialized cells, such as heart or blood cells, in that they can replicate many times, for long periods of time. This ability is what is known as proliferation. Unlike other cells, stem cells also have the ability to differentiate or develop into specialized cells for specific ​organs or to develop into tissues.

In some tissues, such as ​muscle or brain tissue, stem cells can even regenerate to aid in the replacement of damaged cells. Stem cell research attempts to take advantage of the renewal properties of stem cells by utilizing them to generate cells for tissue repair and the treatment of disease.

Where Are Stem Cells Found?

Stem cells come from several sources in the body. The names of the cells below indicate the sources from which they are derived.

Embryonic Stem Cells
These stem cells come from embryos in the early stages of development. They have the ability to differentiate into any type of cell in the initial stages of development and become slightly more specialized as they mature.

Fetal Stem Cells
These stem cells come from a fetus. At about nine weeks, a maturing embryo enters into the fetal stage of development. Fetal stem cells are found in fetal tissues, blood and bone marrow. They have the potential to develop into almost any type of cell.

Umbilical Cord Blood Stem Cells
These stem cells are derived from umbilical cord blood.
Umbilical cord stem cells are similar to those found in mature or adult stem cells. They are specialized cells that develop into specific types of cells.

Placental Stem Cells
These stem cells are contained within the placenta. Like cord blood stem cells, these cells are specialized cells that develop into specific types of cells. Placentas, however, contain several times more stem cells than do umbilical cords.

Adult Stem Cells
These stem cells are present in mature body tissues in infants, children, and adults. They may also be found in fetal and umbilical cord blood cells. Adult stem cells are specific to a particular tissue or organ and produce the cells within that particular tissue or organ. These stem cells help to maintain and repair organs and tissues throughout a person's life.

Types of Stem Cells

Stem cells can be categorized into five types based on their ability to differentiate or their potency. The stem cell types are as follows:

Totipotent Stem Cells
These stem cells have the ability to differentiate into any type of cell in the body. Totipotent stem cells develop during sexual reproduction when male and female gametes fuse during fertilization to form a zygote. The zygote is totipotent because its cells can become any type of cell and they have limitless replicative abilities.
As the zygote continues to divide and mature, its cells develop into more specialized cells called pluripotent stem cells.

Pluripotent Stem Cells
These stem cells have the ability to differentiate into several different types of cells. Specialization in pluripotent stem cells is minimal and therefore they can develop into almost any type of cell. Embryonic stem cells and fetal stem cells are two types of pluripotent cells.

Induced pluripotent stem cells (iPS cells) are genetically altered adult stem cells that are induced or prompted in a laboratory to take on the characteristics of embryonic stem cells. Although iPS cells behave like and express some of the same genes that are expressed normally in embryonic stem cells, they are not exact duplicates of embryonic stem cells.

Multipotent Stem Cells
These stem cells have the ability to differentiate into a limited number of specialized cell types. Multipotent stem cells typically develop into any cell of a particular group or type. For example, bone marrow stem cells can produce any type of blood cell.
However, bone marrow cells don't produce heart cells. Adult stem cells and umbilical cord stem cells are examples of multipotent cells.
Mesenchymal stem cells are multipotent cells of bone marrow that have the ability to differentiate into several types of specialized cells related to, but not including, blood cells. These stem cells give rise to cells that form specialized connective tissues, as well as cells that support the formation of blood.​
Oligopotent Stem Cells
These stem cells have the ability to differentiate into just a few types of cells. A lymphoid stem cell is an example of an oligopotent stem cell. This type of stem cell can not develop into any type of blood cell as bone marrow stem cells can. They only give rise to blood cells of the lymphatic system, such as T cells.
Unipotent Stem Cells
These stem cells have unlimited reproductive capabilities, but can only differentiate into a single type of cell or tissue. Unipotent stem cells are derived from multipotent stem cells and formed in adult tissue. Skin cells are one of the most prolific examples of unipotent stem cells. These cells must readily undergo cell division to replace damaged cells.

Sources: Stem Cell Basics: Introduction. In Stem Cell Information [World Wide Web site]. Bethesda, MD: National Institutes of Health, U.S. Department of Health and Human Services, 2002. Available at (http://stemcells.nih.gov/info/basics/pages/basics1.aspx) ​

Where Are Stem Cells Found In The Body?


Where are stem cells found in the body?


Stem cells come from different places, depending on the type of stem cell you are talking about.

In leukemia and lymphoma, stem cells may be transplanted into the person with cancer to help replenish the bone marrow for production of new blood cells. In these cases, the stem cells come from the bone marrow or bloodstream of a living donor. Stem cells of this type are also known as adult stem cells and, more specifically, hematopoietic stem cells, since they give rise to blood cells.


The stem cell transplants performed in people with cancer do not use any embryonic cells.When it comes to stem cell research, however, stem cells might come from any number of different sources, including from specialized tissues of living human donors or sacrificed human embryos.

Stem Cells for Transplants (HSCT) in Cancer
The bone marrow makes all of your blood cells: red cells, white cells, and other cells called megakaryocytes that make platelets to help with clotting. Hematopoietic stem cells within the bone marrow are the “parents” or progenitors of all of these different cell types that form the blood.

These hematopoietic stem cells are transplanted into a person with cancer to help replenish healthy bone marrow and blood cells; high doses of chemotherapy kill cancer cells effectively, but also kill stem cells in the bone marrow. So, shortly after chemotherapy, stem cells are given into a vein, much like a blood transfusion.
The cells take some time to settle into the bone marrow and begin to grow and make new healthy blood cells.
Bone Marrow vs. Peripheral Blood

Years ago, the only source for stem cells used in transplants was bone marrow. Then a small number of blood-forming stem cells was discovered circulating out in the peripheral bloodstream.

Doctors learned to collect these stem cells from the circulating blood -- and to use them for transplants. This type of transplant, known as a peripheral blood stem cell transplant, or PBSCT, has become more common than transplants from bone marrow, however, both methods are in use today.

Other Stem Cells

In other cases, and in the absence of additional information, the term "stem cell" is ambiguous as to the source of the cells. Additional descriptors are needed to tell you what type of stem cell it is and where it comes from. There are currently three main categories of stem cells; each one has a different source:
Embryonic stem cells are the most controversial stem cells since they come from human embryos that have been destroyed, or harvested for science.

According to the National Institutes of Health (NIH) stem cells from sacrificed human embryos were first grown in a laboratory in 1998; they were originally “created for reproductive purposes,” but when they were no longer needed, they were donated with informed consent of "the donor" -- presumably the egg donor, but possibly the sperm donor -- or perhaps both -- in any case, consent obviously does not refer to the developing human embryo.

Children are usually deemed unable to provide consent for medical procedures prior to the age 16 years; sometimes 18 years of age is used as the threshold age for consent.

Adult stem cells are also called somatic stem cells. These stem cells are obtained without the destruction of a human embryo. Hematopoietic or blood-forming stem cells are an example of this kind of stem cell, and they have been used for generations. The term “somatic” means “body” and underscores that these cells come from a living body and not from a sacrificed embryo. Scientists have found adult stem cells in many more tissues than once thought possible. According to the NIH, adult stem cells have been found in many organs and tissues, including brain, bone marrow, peripheral blood, blood vessels, skeletal muscle, skin, teeth, heart, gut, liver, ovarian cells, and testis.

Induced pluripotent stem cells, or iPSCs, are adult cells that have been genetically reprogrammed to be more like embryonic stem cells. The source is adult or somatic cells and not embryonic cells, but these are adult cells that have undergone genetic engineering.

According to UCLA's Center for Regenerative Medicine and Stem Cell Research, iPSCs usually start out as skin cells or blood cells, which then undergo genetic programming. It is currently possible to produce almost any desired cell type from any patient's cells. Laboratory research has shown promising results using these cells to correct a number of genetic disorders of the blood, nervous system and muscles.
Updated February 2016, TI.

National Institutes of Health. Stem Cell Information. Stem Cell Basics
Simara P, Motl JA, Kaufman DS. Pluripotent Stem Cells and Gene Therapy. Transl Res. 2013;161(4):284-292.
Al-Shamekh S, Goldberg J. Retinal Repair with Induced Pluripotent Stem Cells. Transl Res. 2014;163(4):377-386.
Finkbeiner SR, Spence JR. A Gutsy Task: Generating Intestinal Tissue from Human Pluripotent Stem Cells. Digestive Diseases and Sciences. 2013;58(5):1176-1184.
Priori SG, Napolitano C, Di Pasquale E, Condorelli G. Induced pluripotent stem cell–derived cardiomyocytes in studies of inherited arrhythmias. J Clin Invest. 2013;123(1):84-91.
UCLA. Induced Pluripotent Stem Cells (iPS)


What are adult stem cells?

An adult stem cell is thought to be an undifferentiated cell, found among differentiated cells in a tissue or organ.  The adult stem cell can renew itself and can differentiate to yield some or all of the major specialized cell types of the tissue or organ. The primary roles of adult stem cells in a living organism are to maintain and repair the tissue in which they are found. Scientists also use the term somatic stem cell instead of adult stem cell, where somatic refers to cells of the body (not the germ cells, sperm or eggs). Unlike embryonic stem cells, which are defined by their origin (cells from the preimplantation-stage embryo), the origin of adult stem cells in some mature tissues is still under investigation.
Research on adult stem cells has generated a great deal of excitement. Scientists have found adult stem cells in many more tissues than they once thought possible. This finding has led researchers and clinicians to ask whether adult stem cells could be used for transplants. In fact, adult hematopoietic, or blood-forming, stem cells from bone marrow have been used in transplants for more than 40 years. Scientists now have evidence that stem cells exist in the brain and the heart, two locations where adult stem cells were not at first expected to reside. If the differentiation of adult stem cells can be controlled in the laboratory, these cells may become the basis of transplantation-based therapies.
The history of research on adult stem cells began more than 60 years ago. In the 1950s, researchers discovered that the bone marrow contains at least two kinds of stem cells. One population, called hematopoietic stem cells, forms all the types of blood cells in the body. A second population, called bone marrow stromal stem cells (also called mesenchymal stem cells, or skeletal stem cells by some), were discovered a few years later. These non-hematopoietic stem cells make up a small proportion of the stromal cell population in the bone marrow and can generate bone, cartilage, and fat cells that support the formation of blood and fibrous connective tissue.
In the 1960s, scientists who were studying rats discovered two regions of the brain that contained dividing cells that ultimately become nerve cells. Despite these reports, most scientists believed that the adult brain could not generate new nerve cells. It was not until the 1990s that scientists agreed that the adult brain does contain stem cells that are able to generate the brain's three major cell types—astrocytes and oligodendrocytes, which are non-neuronal cells, and neurons, or nerve cells.

A. Where are adult stem cells found, and what do they normally do?

Adult stem cells have been identified in many organs and tissues, including brain, bone marrow, peripheral blood, blood vessels, skeletal muscle, skin, teeth, heart, gut, liver, ovarian epithelium, and testis. They are thought to reside in a specific area of each tissue (called a "stem cell niche"). In many tissues, current evidence suggests that some types of stem cells are pericytes, cells that compose the outermost layer of small blood vessels. Stem cells may remain quiescent (non-dividing) for long periods of time until they are activated by a normal need for more cells to maintain tissues, or by disease or tissue injury.
Typically, there is a very small number of stem cells in each tissue and, once removed from the body, their capacity to divide is limited, making generation of large quantities of stem cells difficult. Scientists in many laboratories are trying to find better ways to grow large quantities of adult stem cells in cell culture and to manipulate them to generate specific cell types so they can be used to treat injury or disease. Some examples of potential treatments include regenerating bone using cells derived from bone marrow stroma, developing insulin-producing cells for type 1 diabetes, and repairing damaged heart muscle following a heart attack with cardiac muscle cells.

B. What tests are used to identify adult stem cells?

Scientists often use one or more of the following methods to identify adult stem cells: (1) label the cells in a living tissue with molecular markers and then determine the specialized cell types they generate; (2) remove the cells from a living animal, label them in cell culture, and transplant them back into another animal to determine whether the cells replace (or "repopulate") their tissue of origin.
Importantly, scientists must demonstrate that a single adult stem cell can generate a line of genetically identical cells that then gives rise to all the appropriate differentiated cell types of the tissue. To confirm experimentally that a putative adult stem cell is indeed a stem cell, scientists tend to show either that the cell can give rise to these genetically identical cells in culture, and/or that a purified population of these candidate stem cells can repopulate or reform the tissue after transplant into an animal.

C. What is known about adult stem cell differentiation?

As indicated above, scientists have reported that adult stem cells occur in many tissues and that they enter normal differentiation pathways to form the specialized cell types of the tissue in which they reside.
Normal differentiation pathways of adult stem cells. In a living animal, adult stem cells are available to divide for a long period, when needed, and can give rise to mature cell types that have characteristic shapes and specialized structures and functions of a particular tissue. The following are examples of differentiation pathways of adult stem cells (Figure 2) that have been demonstrated in vitro or in vivo.
“Hematopoietic and stromal cell differentiation.” The figure shows a long bone, with marrow in its center and an enlargement of the bone/marrow interface in a boxed inset, with cell types identified.  Cell types shown include the osteocytes embedded in the noncellular bone matrix, the osteoclast, pericytes around tiny blood vessels, adipocytes, and stromal cells.  Using arrows, the artist has drawn illustrations of the lineages of marrow and stromal cells. Marrow lineage:  a hematopoietic stem cell gives rise to a multipotent stem cell, which can divide to produce one of two possible cell types:  (1) a myeloid progenitor cell, which is capable of producing neutrophils, basophils, eosinophils, monocytes/macrophages, platelets, and red blood cells or (2) a lymphoid progenitor cell, which gives rise to natural killer (NK) cells, T lymphocytes, and B lymphocytes. Stromal lineage:  a stromal stem cell gives rise to bone cells, including pre-osteoblasts, osteoblasts, lining cells, and osteocytes. The artist has also indicated two other cell types that the bone marrow may be capable of producing:  skeletal muscle stem cells, and hepatocyte stem cells.  Each possible lineage is followed by a question mark, to indicate that scientists do not agree whether or not bone marrow is capable of producing these two cell types.
Figure 2. Hematopoietic and stromal stem cell differentiation. Click here for larger image. (© 2008 Terese Winslow)
  • Hematopoietic stem cells give rise to all the types of blood cells: red blood cells, B lymphocytes, T lymphocytes, natural killer cells, neutrophils, basophils, eosinophils, monocytes, and macrophages.
  • Mesenchymal stem cells have been reported to be present in many tissues. Those from bone marrow (bone marrow stromal stem cells, skeletal stem cells) give rise to a variety of cell types: bone cells (osteoblasts and osteocytes), cartilage cells (chondrocytes), fat cells (adipocytes), and stromal cells that support blood formation. However, it is not yet clear how similar or dissimilar mesenchymal cells derived from non-bone marrow sources are to those from bone marrow stroma.
  • Neural stem cells in the brain give rise to its three major cell types: nerve cells (neurons) and two categories of non-neuronal cells—astrocytes and oligodendrocytes.
  • Epithelial stem cells in the lining of the digestive tract occur in deep crypts and give rise to several cell types: absorptive cells, goblet cells, Paneth cells, and enteroendocrine cells.
  • Skin stem cells occur in the basal layer of the epidermis and at the base of hair follicles. The epidermal stem cells give rise to keratinocytes, which migrate to the surface of the skin and form a protective layer. The follicular stem cells can give rise to both the hair follicle and to the epidermis.
Transdifferentiation. A number of experiments have reported that certain adult stem cell types can differentiate into cell types seen in organs or tissues other than those expected from the cells' predicted lineage (i.e., brain stem cells that differentiate into blood cells or blood-forming cells that differentiate into cardiac muscle cells, and so forth). This reported phenomenon is called transdifferentiation.
Although isolated instances of transdifferentiation have been observed in some vertebrate species, whether this phenomenon actually occurs in humans is under debate by the scientific community. Instead of transdifferentiation, the observed instances may involve fusion of a donor cell with a recipient cell. Another possibility is that transplanted stem cells are secreting factors that encourage the recipient's own stem cells to begin the repair process. Even when transdifferentiation has been detected, only a very small percentage of cells undergo the process.
In a variation of transdifferentiation experiments, scientists have recently demonstrated that certain adult cell types can be "reprogrammed" into other cell types in vivo using a well-controlled process of genetic modification (see Section VI for a discussion of the principles of reprogramming). This strategy may offer a way to reprogram available cells into other cell types that have been lost or damaged due to disease. For example, one recent experiment shows how pancreatic beta cells, the insulin-producing cells that are lost or damaged in diabetes, could possibly be created by reprogramming other pancreatic cells. By "re-starting" expression of three critical beta cell genes in differentiated adult pancreatic exocrine cells, researchers were able to create beta cell-like cells that can secrete insulin. The reprogrammed cells were similar to beta cells in appearance, size, and shape; expressed genes characteristic of beta cells; and were able to partially restore blood sugar regulation in mice whose own beta cells had been chemically destroyed. While not transdifferentiation by definition, this method for reprogramming adult cells may be used as a model for directly reprogramming other adult cell types.
In addition to reprogramming cells to become a specific cell type, it is now possible to reprogram adult somatic cells to become like embryonic stem cells (induced pluripotent stem cells, iPSCs) through the introduction of embryonic genes. Thus, a source of cells can be generated that are specific to the donor, thereby increasing the chance of compatibility if such cells were to be used for tissue regeneration. However, like embryonic stem cells, determination of the methods by which iPSCs can be completely and reproducibly committed to appropriate cell lineages is still under investigation.

D. What are the key questions about adult stem cells?

Many important questions about adult stem cells remain to be answered. They include:
  • How many kinds of adult stem cells exist, and in which tissues do they exist?
  • How do adult stem cells evolve during development and how are they maintained in the adult? Are they "leftover" embryonic stem cells, or do they arise in some other way?
  • Why do stem cells remain in an undifferentiated state when all the cells around them have differentiated? What are the characteristics of their “niche” that controls their behavior?
  • Do adult stem cells have the capacity to transdifferentiate, and is it possible to control this process to improve its reliability and efficiency?
  • If the beneficial effect of adult stem cell transplantation is a trophic effect, what are the mechanisms? Is donor cell-recipient cell contact required, secretion of factors by the donor cell, or both?
  • What are the factors that control adult stem cell proliferation and differentiation?
  • What are the factors that stimulate stem cells to relocate to sites of injury or damage, and how can this process be enhanced for better healing?

Stem Cell Year-in-Review 2016

Stem Cell Year-in-Review 2016
If we picked one word to define the past year in the stem cell field, it would have to be ‘therapy.’

While many important developments impacted the field, two that garnered significant public, political and scientific attention in 2016 were the proliferation of clinics using unproven stem cell “therapies,” and the steps forward in therapeutic modification of human oocytes (unfertilized eggs) through a process called mitochondrial replacement therapy (MRT).

Within the stem cell field, unproven stem cell therapies, or “treatments” that lack rigorous scientific proof of their safety and effectiveness, have been a growing concern. Clinics often offer these types of treatments directly to consumers, marketing them online or in recruiting seminars, often at substantial cost. The treatments typically use autologous stem cells (those obtained from one’s body) to treat everything from bad knees to incurable disease such as ALS or Parkinson’s. In some countries, this phenomenon traditionally has been called stem cell tourism because it involved travel to another country with less stringent regulations to obtain treatment. Two publications1,2 in 2016 debunked that concept, showing these clinics are found throughout the world. The U.S. alone has an estimated 570 clinics. That puts the U.S. among the countries with the highest number of such clinics, along with India, Mexico, China, Australia, United Kingdom, Thailand, Malaysia and Germany.

Like many countries, the U.S. struggles to regulate unproven stem cell therapies. In 2016, the Food and Drug Administration (FDA), the branch of government charged with regulating cellular therapies, took action to address these issues, proposing revised guidelines that could change how stem cells, especially autologous stem cells, are regulated. The ISSCR was among the organizations and individual scientists that supported the need for careful oversight and regulation of the therapeutic application of stem cells. The FDA has yet to issue a final ruling, but a Perspective article written by three FDA executives published in the New England Journal of Medicine may provide insight into their thinking.

Another big story in 2016 was the first reported use of MRT in humans. This relatively new technology has shown to be effective in animal models for intercepting mitochondrial disease before it’s passed from mother to child. The approach works by replacing the mother’s mutant mitochondria responsible for disease with healthy mitochondria from a donor. The chimeric oocytes (that contain the mother’s nuclear DNA and the donor’s mitochondria) then undergo fertilization through standard IVF methods.

A U.S. fertility specialist reported the birth of a baby boy from a mother who was a carrier for mitochondrial disease that lead to the death of her previously-born children. The MRT procedure is not approved in the U.S., so doctors performed it in Mexico, which led to many concerns within and outside the scientific community. Among them, how do regulatory agencies and the scientific community deal with procedures approved in some countries but not others? Whether and how should we advance human health using technology that alters a natural state? How far should science be applied in influencing health-related outcomes?

This concern was also brought to the forefront of the scientific and public consciousness when a report by Chinese scientists described the use of CRISPR-Cas to modify a gene in human embryos making them resistant to HIV infection [to learn more about CRISPR-Cas, read our previous blog].  Although the embryos were not viable and not intended for clinical use, this proof-of-principle research raised ethical concerns and emphasized the need to continue to have discussions on the scientific and social impact of this technology and its use in countries around the world.

While 2016 saw many articles on unproven stem cell therapies, and the controversial use of MRT, there were many positive stories about potential stem cell-derived therapies that are successfully moving into the clinic for testing. Interventions for macular degeneration, stoke, cancers, and sickle cell disease, among others, are in approved clinical trials in California (U.S.), London, Japan, and elsewhere. These trials, unlike the unproven therapies, have been approved by the appropriate regulatory agencies and will be rigorously tested so that the risks and benefits are more fully understood before (hopefully) being made available to patients. We look forward to continued advances in 2017 that support the rigorous, ethical standards set forth in the 2016 Guidelines for Stem Cell Research and Clinical Translation, and help address issues of human health around the world.

Second Chinese team reports gene editing in human embryos

Study used CRISPR technology to introduce HIV-resistance mutation into embryos.

Early-stage human embryos have been edited by scientists.
Researchers in China have reported editing the genes of human embryos to try to make them resistant to HIV infection. Their paper1 — which used CRISPR-editing tools in non-viable embryos that were destroyed after three days — is only the second published claim of gene editing in human embryos.
In April 2015, a different China-based team announced that they had modified a gene linked to a blood disease in human embryos (which were also not viable, and so could not have resulted in a live birth)2. That report — a world first — fuelled global deliberations over the ethics of modifying embryos and human reproductive cells, and led to calls for a moratorium on even such proof-of-principle research.
At the time, rumours swirled that other teams had conducted similar experiments. Sources in China told Nature’s news team that a handful of papers had been submitted for publication. The latest paper, which appeared in the Journal of Assisted Reproduction and Genetics on 6 April, might be one of these. Nature’s news team has asked the paper’s corresponding author, stem-cell scientist Yong Fan, for comment, but had not heard from him by the time of this report.

HIV resistance

In the paper, Fan, who works at Guangzhou Medical University in China, and his team say that they collected a total of 213 fertilized human eggs between April and September 2014. The fertilized eggs, donated by 87 patients, were unsuitable for implantation as part of in vitro fertility therapy, because they contained an extra set of chromosomes.
Fan’s team used CRISPR–Cas9 genome editing to introduce into some of the embryos a mutation that cripples an immune-cell gene called CCR5. Some humans naturally carry this mutation (known as CCR5Δ32) and they are resistant to HIV, because the mutation alters the CCR5 protein in a way that prevents the virus from entering the T cells it tries to infect.
Genetic analysis showed that 4 of 26 human embryos targeted were successfully modified. But not all the embryos’ chromosomes harboured the CCR5Δ32 mutation — some contained unmodified CCR5, whereas others had acquired different mutations.
George Daley, a stem-cell biologist at Children’s Hospital Boston in Massachusetts, says that the paper’s main advance is the use of CRISPR to introduce a precise genetic modification successfully. “This paper doesn’t look like it offers much more than anecdotal evidence that it works in human embryos, which we already knew,” he says. “It’s certainly a long way from realizing the intended potential” — a human embryo with all its copies of CCR5 inactivated.
“It just emphasizes that there are still a lot of technical difficulties to doing precision editing in human embryo cells,” says Xiao-Jiang Li, a neuroscientist at Emory University in Atlanta, Georgia. He thinks that researchers should work out these kinks in non-human primates, for example, before continuing to modify the genomes of human embryos using techniques such as CRISPR.

Ethics of experiments

Tetsuya Ishii, a bioethicist at Hokkaido University in Sapporo, Japan, sees no problem with how the experiments were conducted — a local ethics committee approved them, and the egg donors gave their informed consent — but he questions their necessity. “Introducing CCR5Δ32 and trying repair, even in non-viable embryos, is just playing with human embryos,” Ishii says.
Fan's team writes in the paper that proof-of-principle experiments for human-embryo editing such as theirs are important to conduct while the ethical and legal issues of germline modification are being hashed out. “We believe that any attempt to generate genetically modified humans through the modification of early embryos needs to be strictly prohibited until we can resolve both ethical and scientific issues,” they write.
Daley sees a stark contrast between Fan’s work and research approved in February by UK fertility regulators that will allow CRISPR genome editing of human embryos. Those experiments, led by developmental biologist Kathy Niakan at the Francis Crick Institute in London, will inactivate genes involved in very early embryo development, in hopes of understanding why some pregnancies terminate. (The work will be done in viable embryos, but the researchers' licence requires that experiments be stopped within 14 days.)
Earlier this year, developmental biologist Robin Lovell-Badge, also at the Francis Crick Institute, told Nature that he thought that the carefully considered UK approval might embolden other researchers who are interested in pursuing embryo-editing research. “If they've been doing it in China, we may see several manuscripts begin to appear,” he said.
Whereas Niakan's work is answering questions intrinsic to embryology, Fan's work is establishing proof of principle for what would need to be done to generate an individual with resistance to HIV, Daley adds. “That means the science is going forward before there’s been the general consensus after deliberation that such an approach is medically warranted," he says.

Stem Cell Basics

Stem cells are the foundation from which all parts of the human body grow.

Cells in the human body

The human body comprises more than 200 types of cells, and every one of these cell types arises from the zygote, the single cell that forms when an egg is fertilized by a sperm. Within a few days, that single cell divides over and over again until it forms a blastocyst, a hollow ball of 150 to 200 cells that give rise to every single cell type a human body needs to survive, including the umbilical cord and the placenta that nourishes the developing fetus.

Basic cell biology

Each cell type has its own size and structure appropriate for its job. Skin cells, for example, are small and compact, while nerve cells that enable you to wiggle your toes have long, branching nerve fibers called axons that conduct electrical impulses.
Cells with similar functionality form tissues, and tissues organize to form organs. Each cell has its own job within the tissue in which it is found, and all of the cells in a tissue and organ work together to make sure the organ functions properly.
Human CellRegardless of their size or structure, all human cells start with these things in common:
  • nucleus that contains DNA, the genetic library for the entire body. Different cells read and carry out different instructions from the DNA, depending on what those cells are designed to do. Your DNA determines virtually everything about your body, from the color of your eyes to your blood type and even how susceptible you are to certain diseases. Some diseases and conditions, such as color blindness, also are passed down through DNA.
  • Cytoplasm – the liquid outside the nucleus. The cytoplasm contains various components that make the materials that the cell needs to do its job.
  • Cell DivisionThe cell membrane – the surface of the cell, a complex structure that sends and receives signals from other cells and lets material in and out of the cell. Cells have to be able to communicate to work together in tissues and organs.
Most cells divide. Shortly before division, the DNA replicates and then the cell divides into two daughter cells. Each has a complete copy of the original cell’s DNA, cytoplasm and cell membrane.

About stem cells

Stem cells are the foundation of development in plants, animals and humans. In humans, there are many different types of stem cells that come from different places in the body or are formed at different times in our lives. These include embryonic stem cells that exist only at the earliest stages of development and various types of tissue-specific (or adult) stem cells that appear during fetal development and remain in our bodies throughout life.
Stem cells are defined by two characteristics:
  • They can make copies of themselves, or self-renew
  • They can differentiate, or develop, into more specialized cells
Beyond these two things, though, stem cells differ a great deal in their behaviors and capabilities.
Embryonic stem cells are pluripotent, meaning they can generate all of the body’s cell types but cannot generate support structures like the placenta and umbilical cord.
Other cells are multipotent, meaning they can generate a few different cell types, generally in a specific tissue or organ.
As the body develops and ages, the number and type of stem cells changes. Totipotent cells are no longer present after dividing into the cells that generate the placenta and umbilical cord. Pluripotent cells give rise to the specialized cells that make up the body’s organs and tissues. The stem cells that stay in your body throughout your life are tissue-specific, and there is evidence that these cells change as you age, too – your skin stem cells at age 20 won’t be exactly the same as your skin stem cells at age 80.

Types of Stem Cells

Stem cells

Stem cells are the foundation for every organ and tissue in your body. There are many different types of stem cells that come from different places in the body or are formed at different times in our lives. These include embryonic stem cells that exist only at the earliest stages of development and various types of tissue-specific (or adult) stem cells that appear during fetal development and remain in our bodies throughout life.
All stem cells can self-renew (make copies of themselves) and differentiate (develop into more specialized cells). Beyond these two critical abilities, though, stem cells vary widely in what they can and cannot do and in the circumstances under which they can and cannot do certain things. This is one of the reasons researchers use all types of stem cells in their investigations.
In this section:
  • Embryonic stem cells
  • Tissue-specific stem cells
  • Mesenchymal stem cells
  • Induced pluripotent stem cells

Embryonic stem cells

Embryonic stem cells are obtained from the inner cell mass of the blastocyst, a mainly hollow ball of cells that, in the human, forms three to five days after an egg cell is fertilized by a sperm. A human blastocyst is about the size of the dot above this “i.”
In normal development, the cells inside the inner cell mass will give rise to the more specialized cells that give rise to the entire body—all of our tissues and organs. However, when scientists extract the inner cell mass and grow these cells in special laboratory conditions, they retain the properties of embryonic stem cells.
Embryonic stem cells are pluripotent, meaning they can give rise to every cell type in the fully formed body, but not the placenta and umbilical cord. These cells are incredibly valuable because they provide a renewable resource for studying normal development and disease, and for testing drugs and other therapies. Human embryonic stem cells have been derived primarily from blastocysts created by in vitro fertilization (IVF) for assisted reproduction that were no longer needed.

Tissue-specific stem cells

Tissue-specific stem cells (also referred to as somatic or adult stem cells) are more specialized than embryonic stem cells. Typically, these stem cells can generate different cell types for the specific tissue or organ in which they live.
For example, blood-forming (or hematopoietic) stem cells in the bone marrow can give rise to red blood cells, white blood cells and platelets. However, blood-forming stem cells don’t generate liver or lung or brain cells, and stem cells in other tissues and organs don’t generate red or white blood cells or platelets.
Some tissues and organs within your body contain small caches of tissue-specific stem cells whose job it is to replace cells from that tissue that are lost in normal day-to-day living or in injury, such as those in your skin, blood, and the lining of your gut.
Tissue-specific stem cells can be difficult to find in the human body, and they don’t seem to self-renew in culture as easily as embryonic stem cells do. However, study of these cells has increased our general knowledge about normal development, what changes in aging, and what happens with injury and disease.

Mesenchymal Stem Cells:
You may hear the term “mesenchymal stem cell” or MSC to refer to cells isolated from stroma, the connective tissue that surrounds other tissues and organs. Cells by this name are more accurately called “stromal cells” by many scientists. The first MSCs were discovered in the bone marrow and were shown to be capable of making bone, cartilage and fat cells. Since then, they have been grown from other tissues, such as fat and cord blood. Various MSCs are thought to have stem cell, and even immunomodulatory, properties and are being tested as treatments for a great many disorders, but there is little evidence to date that they are beneficial. Scientists do not fully understand whether these cells are actually stem cells or what types of cells they are capable of generating. They do agree that not all MSCs are the same, and that their characteristics depend on where in the body they come from and how they are isolated and grown.

Induced pluripotent stem cells

Induced pluripotent stem (iPS) cells are cells that have been engineered in the lab by converting tissue-specific cells, such as skin cells, into cells that behave like embryonic stem cells. IPS cells are critical tools to help scientists learn more about normal development and disease onset and progression, and they are also useful for developing and testing new drugs and therapies.
While iPS cells share many of the same characteristics of embryonic stem cells, including the ability to give rise to all the cell types in the body, they aren’t exactly the same. Scientists are exploring what these differences are and what they mean. For one thing, the first iPS cells were produced by using viruses to insert extra copies of genes into tissue-specific cells. Researchers are experimenting with many alternative ways to create iPS cells so that they can ultimately be used as a source of cells or tissues for medical treatments.

Stem Cells and Research

Stem cell science informs our understanding of the human body and approach to medicine.

These are just a few of the ways stem cells are being used:
  • To study normal human development. Scientists are investigating how stem cells form tissues and organs, how aging impacts their function and their role in various diseases and conditions. A better understanding of the inner working of living organisms leads to earlier detection, better diagnosis and more effective treatments for diseases and injury.
  • In drug discovery, which is the process by which new drugs are identified for a particular disease. Scientists can use stem cells, or tissues grown from them, to search for new drugs that improve their function or alter the progress of disease, as well as to test how drugs might affect different organs (for example, the liver or the kidneys), or how they might affect different people.
  • For cell replacement. Scientists are exploring how to use stem cells to generate tissue that, when transplanted, will take the place of tissue damaged by disease, aging or injury. For example, transplantation of healthy retinal pigment epithelial cells to the eye to replace those lost in macular degeneration is now being tested in clinical trials.
  • For endogenous, or self, repair. Scientists are also exploring ways to stimulate self-repair, coaxing stem cells in the human body to generate healthy cells to heal damaged tissue from within or to prevent further damage.
Stem cell research holds tremendous promise for medical treatments, but scientists still have much to learn about how stem cells, and the specialized cells they generate, work in the body and their capacity for healing. Learn more about clinical translation, the process through which science becomes medicine.

How Science Becomes Medicine

The process by which science becomes medicine is designed to minimize harm and maximize effectiveness.

There is a well-established path by which scientific discoveries are developed into new medical treatments

Clinical translation is the multi-step process of turning scientific discoveries made in the laboratory into real-world medical treatments. This process involves testing a potential new treatment in a series of experiments to assess its safety and effectiveness. When tested on people in the context of a rigorous clinical trial, many possible new treatments fail to be proven safe and effective.
In this section:
  • Basic research
  • Preclinical research
  • Clinical research and patient protections
  • Approval for use

Basic research

Basic research involves figuring out how living organisms, from the cellular level up to the whole animal or person, work and also what can go wrong in disease or injury. Experiments in the lab are where scientists come to understand and test the scientific principles that underlie important medical discoveries.
Scientists in every field are taught to follow the scientific method, a process designed to acquire new knowledge in an objective manner. The basic steps of the scientific method are:
  1. Ask a question
  2. Background research
  3. Come up with a hypothesis, a proposed explanation for the question
  4. Test the hypothesis in a manner in which you can either prove or disprove the hypothesis
  5. Analyze the results of the testing
  6. Make a conclusion
 Scientific Method
Though the scientific method is often presented as a linear sequence of steps, new information or thinking might cause a scientist to back up and repeat steps in the process. Not all steps take place in every scientific inquiry, and they are not always approached in the same order, thus, the scientific method is best considered as general principles.
A vital part of the research process is replication and external (or peer) review. Scientists open up their methods, results and conclusions to the scrutiny of outside experts, typically through publication in peer-reviewed journals. Other scientists replicate the same experiments to reach the same outcomes. In this manner, scientists regularly collaborate with and build upon the discoveries of their peers.

Preclinical research

Preclinical research builds upon the findings of basic research and the understanding gained of disease. It involves the translation of this scientific knowledge into the development of potential treatments. Scientists study how these new treatments work in animals and may also test new treatments on lab-grown animal tissues or human tissues.  Just because a treatment shows promise in an animal, however, does not mean it will be effective in a human, which is why clinical trials are so important.

Clinical research and patient protections

If the results of preclinical research are promising, it may progress to clinical research, or testing in humans.
Clinical trials start with a small number of people and are focused on testing safety. As the procedures are perfected and the risks evaluated, the number of participants is gradually increased and the effectiveness of the treatment is more closely examined. Learn more about Clinical Trials here.
Sometimes, in attempting new surgical techniques or where the disease or condition is rare, treatments might be tried on just one or two people outside the confines of a clinical trial.
During the clinical trial process, there are a number of checks to protect the rights of patients.
Fundamental to the process are:
  • Monitoring of experimental treatments for patient safety and ethical practice. Before beginning, trials should be carefully reviewed by a group of people who together have broad expertise and experience in research, medicine and ethics. These groups, often called Institutional Review Boards (IRBs) or medical ethics review committees, evaluate a number of factors, including the potential risks weighted against the potential benefits.
  • Oversight by regulatory agencies. National oversight agencies, such as the European Medicines Agency (EMA), the U.S. Food and Drug Administration (FDA) or Japan’s Pharmaceuticals and Medical Devices Agency (PMDA), authorize and monitor the development of new treatments. The nature of regulatory agencies and their responsibilities vary from country to country, but most enforce a code of conduct or guidelines for researchers and clinicians to follow to promote safe and effective medical practice.
If you are thinking about a clinical trial for yourself or a loved one, learn more about things you should consider here.

Approval for use

In many countries, a national agency reviews clinical research for evidence of safety and effectiveness, and then approves medical treatments for use by patients. The manner in which medical treatments are marketed is also regulated to ensure companies do not make health claims related to their products that have not been proven through the trial process.
However, the field of stem cell science is new and rapidly changing, and regulation is still catching up. There may come a time when stem cell treatments are regulated consistently by governments across the world. Until such time, people investigating stem cell treatments need to be aware of what is and isn’t regulated in the countries in which they seek treatment. A lack of regulation does not constitute approval or suggest safety or effectiveness. For example, clinics may offer autologous (from your own body) stem cell treatments, which may not be subject to oversight if the cells are minimally manipulated. However, cells taken from an individual are not necessarily safe for use in or as therapy for that same individual.

Buyers beware

Perhaps you’ve heard of “snake oil,” which comes from the 19th century practice of advertising a single elixir as a remedy for all sorts of ailments without clear evidence of quality or health benefit. Unfortunately, false advertising and exaggerated claims are still a problem, and unproven stem cell treatments are among the miracle cures being sold today. Be aware of these things to know if you are considering a stem cell treatment.



The word hydrolysis (loosening, dissolving, breaking, or splitting by the participating action of water) is used when water becomes involved in the metabolism of other materials. Activities that depend on hydrolysis include the breakdown of proteins into their component amino acids and the breakdown of large fatty particles into smaller fatty acid units. Without water , hydrolysis cannot take place. It follows, then, that the hydrolytic function of water also constitutes the metabolism of water itself. What this means is that water itself needs to be broken down first -- hydrolyzed -- before the body can use the various components in food. This is why we need to supply the human body with water before we eat solid foods. Recommended to drink a glass of water , 30 minutes prior your meal time will help the digestion process.

Now that we are on this point, let me once again give you the figures that stress the importance of water as a supplier of energy, especially for brain functions. 

Figure 8.2 : Energy is measured in kilo joules. One kilo Joule is the amount of energy required to raise the temperature of 1 pound of water 1 degree Fahrenheit. [energy]

MgATP(positive)[600] + H2O (water) = 

ADP(3negative)[1500] / ADPH(2negative)[600] 

+ Mg(2positive)[998] / H(positive)[1168] + H2PO

(positive)[318] / HPO4(2negative)[1251]

One unit of magnesium-ATP from the stockpile of energy at the cell membrane has about 600 units of energy before it is hydrolyzed.  When it is hydrolyzed into its component parts , the total energy  content reaches to about 5,835 units. (These figures are taken from an article published by P. George and co-workers : Biochem Biophys Acta 223, no. 1, 1970.) (ATP (adenosine triphosphate), the main source of energy in cells, must be bound to a magnesium ion in order to be biologically active. What is called ATP is often actually Mg-ATP.[4]  ) (Fig. 1. Views of ATP and related structures.  )  

All the foods that we eat and digest need to be hydrolyzed before the human body can tap into their contents. The benefits they offer the body become available because of the "magical" effect  of water, which breaks the products into their more easily digestible and water-energized components.

Dear honorable reader of Healthy Wealth blog-site, 
 Now you know why water is a nutrient and how it generates energy. Water dissolves all the minerals, proteins, starches, and other water-soluble components and , as blood , carries them around the human body for distribution. Think of blood as seawater that has a few breeds of fish in it -- red cells, white cells, platelets, proteins, and enzymes, all swimming to a destination. The blood serum has almost the same mineral consistency and proportions as seawater. 

The human body is in constant need of water. it is losing water though the lungs when we breathe out. The body is losing water in perspiration from the skin, in urine production, and in daily bowel movements. Water deficit causes constipation. A good gauge for the water needs of the human body is the color of urine. 

1. A well-hydrated person produces colorless  urine --- not counting the color of vitamins or color additives in processed edibles.  

2. A comparatively dehydrated person produces translucent-yellow urine. 

3. A truly dehydrated person produces urine that is orange in color.

4. Also: A well hydrated person is never constipated : a constipated person is truly a dehydrated person! 

In the next posts that follows, the importance of minerals and food components are briefly discussed.  For a more thorough understanding of the importance of nutrition in maintaining health and well-being, I recommend the following book. A Complete Illustrated Guide to Vitamins and Minerals : A Practical Approach to a Healthy Diet and Safe Supplementation by Denise Mortimore, BSc , PhD, DHD :ISBN 0607-2717-1. ) You will enjoy reading this  book.

APPENDIX C : Who’s in Charge? Dr. Ihaleakala Hew Len

Who’s in Charge?

Dr. Ihaleakala Hew Len

Thank you for coming along with me in reading this appendix. I am grateful.

I love Self I-Dentity Ho’oponopono and dear 
Morrnah Nalamaku Simeona, Kahuna Lapa’au, 
who so graciously shared it with me in 
November 1982.

This article is based on thoughts logged in my 2005 notebook.

9 January 2005

Problems can be solved without knowing what 
the heck is going on! Realizing and 
appreciating this is sheer relief and joy for me.

Problem solving, part of the purpose for 
existence, is what Self I-Dentity Ho’oponopono 
is about. To solve problems, two questions must 
be addressed: Who am I? Who’s in charge?

To apprehend the nature of the cosmos begins 
with the insight of Socrates: “Know thyself.”

21 January 2005

Who’s in charge?

Most people, including those in the science 
community, deal with the world as being a 
physical entity. Current research in DNA to 
identify causes and remedies for heart disease, 
cancer, and diabetes is a prime example of this.

The Law of Cause and Effect: Physical Model

Cause --(Effect)
Faulty DNA --( Heart Disease )
Faulty DNA --(Cancer)
Faulty DNA --(Diabetes)
Physical -----(Physical Problems)
Physical -----(Environmental Problems)

The Intellect, the Conscious Mind, believes it is 
the problem solver, that it controls what happens 
and what is experienced.

In his book The User Illusion: Cutting 
Consciousness Down to Size, science journalist 
Tor Norretranders paints a different picture of 
Consciousness. He cites research studies, 
particularly those of Professor Benjamin Libet 
of the University of California at San Francisco, 
that show that decisions are made before 
Consciousness makes them, and that the 
Intellect is not aware of this, believing that it 

Norretranders also cites research that shows that 
the Intellect is only conscious of between 15 and 
20 bits of information per second out of millions 
in reaction below its awareness!

If not the Intellect or Consciousness, then who’s 
in charge?

8 February 2005

Memories replaying dictate what the Subconscious 
Mind experiences.

The Subconscious Mind experiences vicariously, 
mimicking and echoing memories replaying. It 
behaves, sees, feels, and decides exactly as
memories dictate. The Conscious Mind too 
operates, without its awareness, by memories 
replaying. They dictate what it experiences, as 
research studies show.

The Law of Cause and Effect: Self I-Dentity Ho’oponopono

Cause (Effect)

Memories Replaying in the Subconscious Mind 
( Physical—Heart Disease)

Memories Replaying in the Subconscious Mind  

Memories Replaying in the Subconscious Mind 

Memories Replaying in the Subconscious Mind (Physical Problems—the Body)

Memories Replaying in the Subconscious Mind  
(Physical Problems—the World)

The body and the world reside in the 
Subconscious Mind as creations of memories 
replaying, rarely as Inspirations.

23 February 2005

The Subconscious Mind and Conscious Mind, 
comprising the Soul, do not generate their own 
ideas, thoughts, feelings, and actions. As noted 
before, they experience vicariously, through 
memories replaying and Inspirations.

But men may construe things after their fashion
Clean from the purpose of the things themselves.
--William Shakespeare

It is essential to realize that the Soul does not 
generate experiences of its own, that it sees as 
memories see, feels as memories feel, behaves 
as memories behave, and decides as memories 
decide. Or, rarely, it sees, feels, behaves, and 
decides as Inspiration sees, feels, behaves, and 

It is crucial in problem solving to realize that the 
body and the world are not the problems in and 
of themselves but the effects, the consequences, 
of memories replaying in the Subconscious 
Mind! Who’s in charge?

Poor soul, the center of my sinful earth,
[Thrall to] these rebel pow’rs that thee array,
Why dost thou pine within and suffer dearth,
Painting thy outward walls so costly gay?
--William Shakespeare, Sonnet 146

12 March 2005

The Void is the foundation of Self I-Dentity, of 
Mind, of the cosmos. It is the precursor 
state to the infusion of Inspirations from Divine 
Intelligence into the Subconscious Mind. 
(See Figure C.1.)

All that scientists know is the cosmos was
spawned from nothing, and will return to the 
nothing from whence it came. The universe 
begins and ends with zero. -- Charles Seife, 
Zero: The Biography of a Dangerous Idea.

FIGURE C.1 State of VoidI

Self I-Dentity
State of Void

Infinite (Divine Intelligence)

Void :-
--Superconscious Mind

--Conscious Mind

--Subconscious Mind


Memories replaying displace the Void of Self I-
Dentity, precluding the manifestation of 
Inspirations. To remedy this displacement, to 
reestablish Self I-Dentity, memories need to be 
transformed to void through transmutation
by Divine Intelligence.

Clean, erase, erase and find your own Shangri-la. 
Where? Within yourself.
--Morrnah Nalamaku Simeona, Kahuna Lapa’au

Nor stony tower, nor walls of beaten brass,
Nor airless dungeon, nor strong links of iron,
Can be retentive to the strength of spirit.
--William Shakespeare, Playwright

22 March 2005

Existence is a gift from Divine Intelligence. And 
the gift is given for the sole purpose of 
reestablishing Self I-Dentity through problem 
solving. Self I-Dentity Ho’oponopono is 
an updated version of an ancient Hawaiian 
problem solving process of repentance, 
forgiveness, and transmutation.

Do not judge, and you will not be judged. Do not 

condemn, and you will not be condemned. 
Forgive and you will be forgiven.
--Jesus as reported in Luke: 6

Ho’oponopono involves the full participation of 
each of the four members of Self I-Dentity—
Divine Intelligence, Superconscious Mind, 
Conscious Mind, and Subconscious Mind—
working together as a unit of one.
Each member has its unique part and function in 
problem solving memories replaying in the 
Subconscious Mind.

The Superconscious Mind is memory free, 
unaffected by memories replaying in the 
Subconscious Mind. It is always one with 
Divine Intelligence. However Divine 
Intelligence moves, so moves the 
Superconscious Mind.

Self I-Dentity operates by Inspiration and 
memory. Only one of them, either memory or 
Inspiration, can be in command of the 
Subconscious Mind at any given moment. The 
Soul of Self I-Dentity serves only one master at 
a time, usually memory the thorn instead of 
Inspiration the rose.
(See Figure C.2.)
FIGURE C.2 State of Inspiration and State of Memory Replaying

30 April 2005

I am the self consumer of my woes.
--John Clare, poet

Void is the common ground, the equalizer, of all 
Self Identities, both “animate” and “inanimate.” 
It is the indestructible and timeless foundation of 
the entire cosmos, seen and unseen.

We hold these truths to be self-evident, that all 
men [all life forms] are created equal. . . .
--Thomas Jefferson, U.S. Declaration 
of Independence

Memories replaying displace the common ground 
of Self I-Dentity, taking the Soul of Mind away 
from its natural position of Void and Infinite.
Although memories displace the Void, they 
cannot destroy it. How can nothing be destroyed?

A house divided against itself cannot stand.
--Abraham Lincoln

5 May 2005

For Self I-Dentity to be Self I-Dentity moment to 
moment requires incessant Ho’oponopono. Like 
memories, incessant Ho’oponopono can never
go on vacation. Incessant Ho’oponopono can 
never retire. Incessant Ho’oponopono can 
never sleep. Incessant Ho’oponopono can 
never stop as . . .

. . . in your days of gladness bear in mind the 
unknown evil [memories replaying] forging on 
behind!--Geoffrey Chaucer, Canterbury Tales

12 May 2005

The Conscious Mind can initiate the Ho’oponopono 
process to release memories or it can engage 
them with blame and thinking. (See Figure C.3.)
FIGURE C.3 Repentance and Forgiveness

(3) Divine Intelligence

(2) Superconscious Mind

(1)Conscious Mind


Self I-Dentity Ho’oponopono
(Problem Solving)
Repentance and Forgiveness

1. Conscious Mind initiates the Ho’oponopono 

problem solving process, a petition to Divine 
Intelligence to transmute memories to Void. It 
acknowledges that the problem is memories 
replaying in its Subconscious Mind, and that it 
is 100 percent responsible for them. The petition 
moves down from the Conscious Mind into the 
Subconscious Mind. (See Figure C.4.)

2. The down flow of the petition into the 
Subconscious Mind gently stirs memories for 
transmutation. The petition then moves up to
the Superconscious Mind from the Subconscious 

3. The Superconscious Mind reviews the petition, 
making changes as appropriate. Because it is 
always in tune with Divine Intelligence, it has 
the capacity to review and make changes. The 
petition is then sent up to Divine Intelligence for 
final review and consideration.

4. After reviewing the petition sent up by the 
Superconscious Mind, Divine Intelligence sends 
transmuting energy down into the 
Superconscious Mind.

5. Transmuting energy then flows from the 
Superconscious Mind down into the Conscious 

6. And transmuting energy then flows down from 
the Conscious Mind into the Subconscious Mind. 
The transmuting energy first neutralizes 
designated memories. The neutralized energies 
are then released into storage, leaving a Void.

12 June 2005

Thinking and blame are memories replaying 

(see Figure C.2).

The Soul can be inspired by Divine Intelligence 

without knowing what the heck is going on. The 
only requirement for Inspiration, Divine 
creativity, is for Self I-Dentity to be Self I-Dentity. 
To be Self I-Dentity requires incessant 
cleansing of memories.

Memories are constant companions of the

 Subconscious Mind.

They never leave the Subconscious Mind to 

go on vacation. They never leave the 
Subconscious Mind to go into retirement. 
Memories never stop their incessant replaying!

The Man of Law’s Tale

O sudden grief that ever art near neighbour

To worldly bliss! Sprinkled with bitterness

The ends of joy in all our earthly labour!

Grief occupies the goal to which we press.

For your own safety think it is no less,

And in your days of gladness bear in mind

The unknown evil forging on behind!

Geoffrey Chaucer, Canterbury Tales

FIGURE C.4 Transmutation by Divine Intelligence.

(4)Divine Intelligence

(5)Superconscious Mind

(6)Conscious Mind

(memory) --> n --> Vision ; Subconscious Mind

To be done with memories once and for all, they 
must be cleansed to nothing once and for all.

It was in Iowa in 1971 that I fell head over heels 
in love for the second time. Dear M, our 
daughter, was born.

As I watched my wife care for M, I fell deeper 
and deeper in love with both of them. I had two 
wonderful people to love now.

After completing graduate school in Utah that 
summer, my wife and I had a choice to make: 
to go home to Hawaii or to continue graduate 
training in Iowa.

As we began life in the Hawkeye State, two 
hurdles immediately confronted us. First, M had 
never stopped crying after we brought her home 
from the hospital!

Second, the worst winter of the century in Iowa 
set in. Each morning for weeks on end I kicked 
the bottom inside of the front door of our 
apartment and hammered its edges with my 
hands to break the entombing ice on the other side.

Around her first year, bloodstains showed up on 
M’s blankets. Only now as I write this sentence, 
I realize that the constant crying was her reaction 
to the severe skin problem that was diagnosed

I cried many a night as I helplessly watched M in 
fitful sleep scratching herself. Steroid medications 
proved powerless to help her.

By age three, blood seeped continuously from 
cracks in the crooks of M’s elbows and knees. 
Blood wept from cracks around the joints of her 
fingers and toes. Thick mantles of hard skin 
covered the inside of her arms and around her 

One day nine years later, after we had returned to 
Hawaii, I was driving home with M and her 
sister. Suddenly, without conscious forethought, 
I found myself turning the car around and 
heading in the direction of my office in Waikiki.

“Oh, you folks have come to visit me,” Morrnah 
said quietly as the three of us trooped into her 
office. As she shuffled papers on her desk, she 
looked up at M. “Did you want to ask me 
something?” she said softly.

M stretched out both arms, revealing years of pain 
and grief  etched in them up and down like 
Phoenician scrolls. “Okay,” came Morrnah’s reply
, and she closed her eyes.

What was Morrnah doing? The creator of Self I-
Dentity Ho’oponopono was doing Self I-Dentity 
Ho’oponopono. A year later, 13 years of bleeding, 
scarring, pain, grief, and medications had come to
an end.---Self I-Dentity Ho’oponopono student

30 June 2005

The purpose of life is to be Self I-Dentity as 
Divinity created Self I-Dentity in its exact 
likeness, Void and Infinite.

All life experiences are expressions of memories 
replaying and Inspirations. Depression, thinking, 
blame, poverty, hate, resentment, and grief are
“fore-bemoanèd moans,” as Shakespeare noted 
in one of his sonnets.

The Conscious Mind has a choice: It can initiate 
incessant cleansing or it can allow memories to 
replay problems incessantly.

12 December 2005

Consciousness working alone is ignorant of Divine 
Intelligence’s most precious gift: Self I-Dentity. As 
such, it is ignorant of what a problem is. This
ignorance results in ineffectual solving of the 
problem. Poor Soul is left to incessant, needless 
grief for its entire existence. How sad.

The Conscious Mind needs to be awakened to the 
gift of Self I-Dentity, “wealth beyond all 

Self I-Dentity is indestructible and eternal, as is its 
Creator, DivineIntelligence. The consequence of 
ignorance is the false reality of senseless and 
relentless poverty, disease, and war and death 
generation after generation.

24 December 2005

The physical is the expression of memories and 
Inspirations taking place in the Soul of Self I-
Dentity. Change the state of Self I-Dentity and 
the state of the physical world changes.

Who’s in charge—inspirations or memories 
replaying? The choice is in the hands of the 
Conscious Mind.

7 February 2006 (A Leap into 2006)

Here are four Self I-Dentity Ho’oponopono problem 
solving processes that can be applied to reestablish 
Self I-Dentity through voiding memories replaying 
problems in the Subconscious Mind:

1. “I love you.” When Soul experience memories 
replaying problems, say to them mentally or 
silently: “I love you, dear memories. I am grateful 
for the opportunity to free all of you and me.” “I 
love you” can be repeated quietly again and again. 
Memories never go on vacation or retire unless 
you retire them. “I love you” can be used even if 
you are not conscious of problems. For example, 
it can be applied before engaging in any activity 
such as making or answering a telephone call or 
before getting into your car to go somewhere.

Love your enemies, do good to those who hate 
you. -- Jesus Christ as reported in Luke: 6

2. “Thank you.” This process can be used with or 
in place of “I love you.” As with “I love you,” it 
can be repeated mentally again and again.

3. Blue solar water. Drinking lots of water is a 
wonderful problem solving practice, particularly 
if it is blue solar water. Get a blue glass container 
with a nonmetallic cover. Pour tap water into the
container. Place the blue glass container either in 
the sun or under an incandescent lamp 

(not a fluorescent lamp) for at least an hour. After 
the water is solarized, it can be used in several 
ways. Drink it. Cook with it. Rinse with it after a 
bath or shower. Fruits and vegetables love being 
washed in blue solar water! As with “I love you” 
and “Thank you” processes, blue solar water 
voids memories replaying problems in the 
Subconscious Mind. So, drink away!

4. Strawberries and blueberries. These fruits 
void memories. They can be eaten fresh or dried. 
They can be consumed as jams, jellies, and even 
syrup on ice cream!

27 December 2005 (A Leap Back into 2005)

I got the idea a few months back of a “talking” 
glossary of the essential “characters” in Self I-
Dentity Ho’oponopono. You can get acquainted 
with each of them at your leisure.

Self I-Dentity: I am Self I-Dentity. I am composed 
of four elements: Divine Intelligence, 
Superconscious Mind, Conscious Mind, and 
Subconscious Mind. My foundation, Void and 
Infinite, is an exact replication of Divine 

Divine Intelligence: I am Divine Intelligence. I 
am the Infinite. I create Self I-Dentities and 
Inspirations. I transmute memories to Void.

Superconscious Mind: I am Superconscious Mind. 
I oversee the Conscious and Subconscious Minds. I 
review and make appropriate changes in the 
Ho’oponopono petition to Divine Intelligence 
initiated by the Conscious Mind. I am unaffected 
by memories replaying in the Subconscious Mind. 
I am always one with Divine Creator.

Conscious Mind: I am Conscious Mind. I have the 
gift of choice. I can allow incessant memories to 
dictate experience for the Subconscious Mind and 
me or I can initiate the release of them through 
incessant Ho’oponopono. I can petition for 
directions from Divine Intelligence.

Subconscious Mind: I am Subconscious Mind. I am
the storehouse for all of the accumulated memories 
from the beginning of creation. I am the place where 
experiences are experienced as memories replaying 
or as Inspirations. I am the place where the body and 
the world reside as memories replaying and as 
Inspirations. I am the place where problems live as 
memories reacting.

Void: I am Void. I am the foundation of Self I-
Dentity and the Cosmos. I am where Inspirations 
spring forth from Divine Intelligence, the Infinite. 
Memories replaying in the Subconscious Mind 
displace me but do not destroy me, precluding 
the inflow of Inspirations from Divine Intelligence.

Infinite: I am Infinite, Divine Intelligence. 
Inspirations flow like fragile roses from me into 
the Void of Self I-Dentity, easily displaced by the 
thorns of memories.

Inspiration: I am Inspiration. I am a creation of 
the Infinite, of Divine Intelligence. I manifest 
from the Void into the Subconscious Mind. I am 
experienced as a brand-new occurrence.

Memory: I am memory. I am a record in the 
Subconscious Mind of a past experience. When 
triggered, I replay past experiences.

Problem: I am problem. I am a memory replaying 
a past experience again in the Subconscious Mind.

Experience: I am experience. I am the effect of 
memories replaying or Inspirations in the 
Subconscious Mind.

Operating System: I am the operating system. I 
operate Self I-Dentity with Void, Inspiration, and 

Ho’oponopono: I am Ho’oponopono. I am an 
ancient Hawaiian problem solving process 
updated for today’s use by Morrnah Nalamaku 
Simeona, Kahuna Lapa’au, recognized as a 
Living Treasure of Hawaii in 1983. I am 
composed of three elements: repentance, 
forgiveness, and transmutation. I am a petition
initiated by the Conscious Mind to Divine 
Intelligence to void memories to reestablish Self 
I-Dentity. I begin in the Conscious Mind.

Repentance: I am repentance. I am the beginning 
of the Ho’oponopono process initiated by the 
Conscious Mind as a petition to Divine 
Intelligence to transmute memories to Void. With 
me, the Conscious Mind acknowledges its 
responsibility for the memories replaying 
problems in its Subconscious Mind, having 
created, accepted, and accumulated them.

Forgiveness: I am forgiveness. Along with 
Repentance, I am a petition from the Conscious 
Mind to Divine Creator to transform memories 
in the Subconscious Mind to Void. Not only is 
the Conscious Mind sorrowful, it is also asking
Divine Intelligence for forgiveness.

Transmutation: I am transmutation. Divine 
Intelligence uses me to neutralize and to release 
memories to Void in the Subconscious Mind. I 
am available for use only by Divine Intelligence.
Just say "I love you."

Wealth: I am wealth. I am Self I-Dentity.

Poverty: I am poverty. I am memories replacing. I 
displace Self I-Dentity, precluding the infusion of 
Inspirations from Divine Intelligence into the 
Subconscious Mind!

Before bringing this visit with you to an end, I 
would like to mention that reading this appendix 
satisfies the prerequisite of attending a day lecture 
if you are considering taking a Self I-Dentity 
Ho’oponopono weekend class.

I wish you peace beyond all understanding.

O Ka Maluhia no me oe.

Peace be with you,

Ihaleakala Hew Len, PhD

Chairman Emeritus

The Foundation of I, Inc. Freedom of the Cosmos