Stephen Holt MD, PhD, DSc, LLD(Hon.) DNM, ChB, FRCP (C), MRCP (UK), FACP, FACG, FACN, FACAM, KSJ, Distinguished Professor of Medicine (Emerite), Scientific Advisor, Natural Clinician LLC, Little Falls, NJ

Many different types of stem cells are being used in research and clinical practice, on a global basis1. Examples of these stem cells include, Human Embryonic Stem Cells (ESC), Human Cord Blood or Placental Stem Cells, Adult Stem Cells (ASC) and even animal stem cells (live cell therapy). While there are clear and obvious advantages of the use of ESC as a consequence of their totipotential nature, the use of such cells in general therapeutics presents insurmountable ethical and moral problems2,3. Although the use of human umbilical stem cells tends to overcome ethical problems, work in this field poses special technical challenges1. These circumstances have led to major interest in the use of ASC. In ASC procedures stem cells are harvested, grown, manipulated and reintroduced1. The overall objective of ASC treatments is to implant ASC which are often coaxed down a pathway of differentiation toward a specific adult somatic cell type in order to replace diseased or ailing tissues.
Adult Stem Cells are pluripotent and they have different degrees of versatility in their ability to engraft and replace damaged tissues. While some ASC appear limited in their ability to differentiate into different cell types, many recent studies show that such cells (especially stromal bone marrow or adipose tissue-derived) may have great versatility, in some circumstances1,4-22. Elegant biotechnology research is being undertaken to transform harvested ASC into highly specialized “functional” cell types for the potential treatment of disorders such as Parkinson’s disease1.
There has been a great deal of high quality research in the application of ASC treatments, especially in the field of human bone marrow transplantation. However, there have been reports of poorly performed ASC treatment in off shore locations which are alleged to be poorly equipped or devoid of important ancillary services to make ASC treatments safe and effective. Recent revisions to U.S. laws that govern stem cell treatments (Obama legislation) have produced widespread interest in the development of stem cell treatment facilities, especially in centers of healthcare excellence. While stem cell therapies are advancing in a meteoric manner, all current “classic” stem cell treatments provide a series of disadvantages or limitations1.
Innovative scientists have been working on the potential use of mobilized in-situ ASC, as a non invasive form of stem cell treatment23,24. This is the process of Induction of Adult Stem Cell Recruitment (IASCR)25. A decade of research has led to current proposals that endogenous or in situ ASC (most notably bone marrow stem cells) can be mobilized from their niches in the body, with the result that they may migrate to various organs and engage in tissue repair or regeneration23-25. While somewhat futuristic, I have proposed that there may be several means whereby endogenous ASC could be released and promoted to differentiate into desired cell types to treat specific organ damage25. This proposal is supported by major advances in the characterization compounds that can induce stem cell differentiation to specific cell types in-vitro1.
The proposed, non invasive technology of IASCR has obvious advantages over the processes of harvesting and reintroduction of ASC. These processes form part of current complex treatment programs1. A body of evidence has emerged that several pharmaceuticals or natural substances are capable of mobilizing ASC from human bone marrow deposits, but some uncertainty surrounds the ability of mobilized ASC to home into damaged tissue and undertake a process of recruitment that will produce the desired treatment outcome of tissue repair or regeneration. However, diseased or damaged tissues provide complex signals to attract regenerative stem cells and the body has a built in homing system that it utilizes when ASC are part of an “internal repair kit”1, 23-25. The development of IASCR represents a new horizon in stem cell treatments that could make stem cell therapies more portable and cost effective25.
The objective of this article is to highlight the ability of combinations of nutrients, botanicals or herbals that can play a role in the mobilization of ASC and their antioxidant protection during their migration. Earlier work is very important in the current proposals and this research must be acknowledged and applauded23, 24. I have called the process described in this article “IASCR” (Induction of Adult Stem Cell Recruitment), with the “R” making a certain assumption of ASC Recruitment, justified or otherwise. With further research, the “R” may stand for Regeneration or Repair of diseased or ailing tissues.

Adult Stem Cells at Work
Adult Stem Cells are ubiquitous in the body and they live in “niches” in many organs1. Most research has been performed with ASC of bone marrow origin, where stem cells are encouraged to proliferate to support the presence of blood components, often following marrow ablation. It has been stated that the very presence of ASC in adults poses questions concerning the exact definition of a stem cell1. While scientists have no problem in discussing the potential of several stem or progenitor cells to form new cell types or engage in tissue renewal, the concept of “stemness” emerges (CT Scott, 2006)1. In brief, “stemness” is the ability of a stem cell to produce different cell types and their ability to engage in self renewal1.
There is a large body of clinical and scientific literature that demonstrates the pluripotential of bone marrow ASC4-22. Bone marrow ASC have been harvested and reinjected into patients, following varying degrees of laboratory manipulation, in order to treat the consequences of degenerative disease. Variable success is apparent in some anecdotal reports on the internet (key search words: adult stem cells). The results of many of these studies are reported in animal experiments in great detail, but many human experiences remain quite anecdotal in their descriptions of clinical outcome25. Bone marrow ASC are engaged in the long term replenishment of all blood elements, but they are composed of a group of non-hematopoietic ASC (stromal cells) which are precursors of bone, cartilage and skeletal tissues24,25. This limited view of ASC has been overturned by many observations of the ability of these ASC to form a much wider range of specific cell types that may play a pivotal role in tissue healing and cellular replacement or regeneration4-22. In simple terms, ASC in a variety of niches in the body could migrate and translocate to a site of tissue damage where they may undergo cellular differentiation that improves organ structure and function1, 4-22.
In their classic article in Medical Hypothesis (2002), Jensen and Drapeau23 describe a hypothesis to support some of the components of what I have termed IASCR. In brief, these scientists highlight the importance of the ability of ASC to target and grow at locations of tissue damage and an ability for this migration to occur after induced mobilization of ASC, most notably from bone marrow. There are other steps to be applied to the concept of IASCR which may include the use of agents to protect stem cells or improve their functionality and enhance their ability to differentiate into the desired types of adult somatic cells25.
Many authors confirm the pluripotent properties and ability of ASC to migrate within the body1,4-25. Such studies include the ability of bone marrow ASC to become functional myocytes, hepatocytes, osteocytes and cells of the central nervous system4-24. Available scientific information permits a clear conclusion that ASC have an ability to migrate from their tissues sites of origin and undergo cellular differentiation that results in a variable degree of repair of many different tissues1.
The laboratory identification of ASC involves a check for the presence of three characteristics that are hallmarks of “stemness” (CT Scott, 2006)1. First, the cells must be able to renew themselves. Second, the cells must have an ability to differentiate into specific cell types and, third, the cells must be transplantable with functional engraftment1. A characteristic of all stem cells is the presence of telomerase, an important marker found in cancer cells1. The presence of telomerase has led to proposals that many types of malignancies originate in stem cells, as a consequence of disorganized cell division.
The residual argument that promoting ASC activity in humans could lead to the development of cancer is not supported by current scientific knowledge or experimentation. At present, it appears to be a reasonable and safe proposal that ASC may be mobilized in the human body without significant adverse effects25. The use of adult stem cell technology has been perceived as widely acceptable in medical practice, because it rests on the relative safety and effectiveness of human bone marrow transplantation1, but regulatory issues concerning the use of adult stem cell therapy in the US remain to be defined with clarity.

Mobilizing ASC
Several drugs, biological agents and nutritional factors exert effects on the mobilization and disposition of otherwise quiescent adult stem cells (Table 1)23-25. A number of nutritional co-factors exert effects on supporting the differentiation of stem cells (e.g. hematopoietic bone marrow stem cells require vitamin D, B12, folic acid and iron for maturation). Recent studies imply that single or combination formulations of natural substances may promote the mobilization of stem cells23-25. Such natural agents include, but may not be limited to: carnosine, blueberries (and other anthocyanidins containing botanicals), green tea derivatives and components of algae (fucoidans and pigments)23-25 (Table 1).

DRUGS (or isolated biologicals in clinical use or trials): IL1, IL3, IL6, Stem Cell Factor(s), erythropoietin, G-CSF etc.
NUTRACEUTICALS: oleic acid, linolenic acid, blueberry, blue-green algae (AFA), green tea, fucoidan and vitamin D3. Putative releasers or antioxidant protection may occur with fucoxanthin, beet root, spirulina, spinach, Ashwagandha, grape seed extract. Cofactors: Vitamin B12, Folate etc.

Table 1. Factors that mobilize Adult Stem Cells (ASC) or provide nutritional support, including antioxidant protection for stem or progenitor cells. The effects of many agents on cell signaling cascades remain underexplored.

In conventional medical practice, a number of drugs have been utilized to stimulate bone marrow stem cell activity (e.g. Granulocyte-Macrophage Colony-Stimulating Factor, GM-CSF or erythropoietin)26, 27. In addition, various combinations of cytokines can facilitate the growth of stem cells in vitro (e.g. IL-1, IL-3, IL-6, erythropoietin and stem cell factors). In vitro comparisons of specific synergistic formulae of putative stem cell supporting nutrients with the actions of the drug GM-CSF imply that such combinations of nutrients may, in some circumstances, increase bone marrow stem cell proliferation to a degree greater than the drug GM-CSF24. Whether or not mobilized adult stem cells can be recruited in a consistent manner and result in engraftment to replace diseased or ailing tissues requires further investigation; and this subject has attracted legal controversies with regulatory authorities or “bounty hunters” who impact the sale of dietary supplements28, 29.
There have been many anecdotal reports of benefit following the use of nutritional agents to support adult stem cell structure and function, but these reports are largely in testimonial format, often displayed on the internet in multilevel marketing information (network selling). However, it is recognized that adult stem cells are often “tissue specific” in their homing characteristics1, 17, 19, 22-24. Moreover, diseased or degenerating cells produce a variety of chemical messengers (e.g. cytokines) or have alterations of cell surface receptors that may attract reparative adult stem cells1, 17, 19. A key, but sometimes overlooked, factor in all adult stem cell technology is to protect the utilized stem cells (in-situ or exogenous allograft administration) from oxidative stress by the use of REDOX balanced antioxidants. Such antioxidants may be present present in several nutritional factors that promote stem cell structure and function25 (e.g. fucoxanthin in fucoidan-containing brown algae30).

Key Contemporary Research
Several scientists have presented experimental or clinical information on the use of specific nutrients or botanicals for the mobilization of ASC23-25. There has been considerable examination and controversy concerning the use of Aphanizomenon flos aquae (AFA or Blue Green Algae) for use as an enhancer of stem cell mobilization28, 29, among other things. In February 2003 a California court ruled that many statements used in the marketing of a branded form of blue green algae were deceptive28, 29. The court ruled that the use of this ingredient was associated with a series of advertising claims that were untrue, unfounded or likely to mislead, to variable degrees28, 29. However, more recent research would seem to imply that there is a scientific basis for the use of blue green algae (AFA) in the nutritional support of stem cell mobilization31.
In well conducted experiments, preparations of AFA containing a novel cyanobacterial ligand for human L-Selectin was shown to have an important role in stem cell biology31. An extract of AFA, enriched for a novel ligand for human L-Selectin was studied in laboratory and human experiments31. L-Selectin is an example of a cell adhesion molecule that plays a role in the retention and mobilization of bone marrow stem cells into the systemic circulation. In brief, scientists showed that oral administration of this extract of AFA contained an L-Selectin blocker which promotes the release of stem cells from the bone marrow by interfering with the functions of CXCR4 cemokine receptors that are specific for stromal derived factor-1 (SDF-1)31.
Any compound that interferes with CXCR4 or SDF-1 will promote stem cell mobilization. These experiments involving the administration of the AFA extract resulted in a 25% increase in the number of circulating stem cells at one hour post administration, in humans, compared with placebo31. Stem cells were measured in blood samples by defining the presence of the CD34+ cell31. These results were found to be reproducible in humans. The scientists commented that the mobilization of stem cells treated by an L-Selectin blocker appears to be of a lower magnitude and greater transience of effects than can be achieved by the administration of Granulocyte Colony Stimulating Factor (G-CSF)31. In separate experiments, scientists have proposed that an enhanced level of circulating CD34+ stem cells a reasonable indicator of good health31.
Several nutrients appear to have an effect on mobilizing bone marrow ASC or stromal cells. For example, vitamin D and 1, 25 dihydroxyvitamin D3 are modulators of several immune functions, including an ability to stimulate the production of progenitor cells32. In addition, oleic and linoleic acids have been shown to promote the proliferation of intestinal stem cells and hematopoietic precursor cells, respectively33, 34. Combinations of these simple nutrients form a significant component of any product used for stem cell support; and they are likely to act in a safe and synergistic manner with specific ASC-mobilizers, such as AFA25. The power of synergy in the use of combinations of nutrients and botanicals has been highlighted in recent studies by Bickford et al (2006)24.
In one important study, Dr. Bickford et al24 undertook laboratory (in-vitro) studies to examine the ability of certain natural substances (nutraceuticals) to work together in a synergistic manner to promote the proliferation of human adult stem cells. In these studies, the scientists directed their attention to hematopoietic (bone marrow) stem cells, with the knowledge that these types of adult stem cells are those that are used routinely in the relatively common practice of human bone marrow transplantation. These studies show that certain nutrients or botanical extracts were able to stimulate bone marrow cell proliferation in a manner that was dose-dependent, i.e. related to the amount of the administered natural substances24.
The experiments by Dr. Bickford et al24 were performed using a positive control substance which is known to stimulate bone marrow stem cell activity. The positive control substance used in these experiments was the drug Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF)26. This pharmaceutical (GM-CSF) is often prescribed by cancer therapists in a routine manner to counteract side effects of toxic drugs that are used in cancer treatments26. Chemotherapeutic drugs often result in bone marrow damage and suppression of immune function.
By using GM-CSF as a control, the scientists were able to make laboratory assessments of the ability of various combinations of nutraceuticals (nutrients or botanicals) to induce positive effects on the stimulation and mobilization of bone marrow stem cells24. The positive control, using GM-CSF, resulted in an anticipated stimulation of bone marrow cell production by a factor of about 46%24. While this ability to stimulate bone marrow cell proliferation was more substantial than the use of any single nutrient or natural substance under investigation, the combined use of natural substances (nutraceuticals) resulted in a greater increase in bone marrow cell proliferation than was observed with GM-CSF24. In other words, in these experiments, combinations of nutrients and botanicals (natural agents) worked better than the control drug (GM-CSF)24.
It is relevant to note that several nutraceuticals that promote adult stem cell production have a benefit of antioxidant actions which may assist in preventing damaged or circulating ASC from oxidative stress (free radical generation)25. Oxidative stress can attack the viability and impair the potentially beneficial functions of “stimulated” or “mobilized” adult stem cells25.
It is quite valuable to examine in detail just how powerful the synergistic effects of nutrient and botanical combinations actually are on bone marrow cell proliferation. Dr. Bickford et al24 showed that a combination of blueberry and green tea extracts caused specific types of bone marrow cell proliferation by a factor of up to 70%; and a simple combination of blueberry extract with vitamin D3 caused an increase of about 62%. When blueberry extract was combined with carnosine, an increase in proliferation of bone marrow cells was observed to be about 83%24.
The experiments of Dr. Bickford et al24 were further extended to the study of various natural substances (nutraceuticals) on the stimulation of stem cells in the bone marrow24. Adult stem cells can be identified and segregated by identification of surface antigens (laboratory markers). A good example of the presence of such surface antigens is the identification of CD 133+ or CD 34+ antigen-receptor expressions. In the experiments, the drug GM-CSF increased CD 34+ and CD 133+ (early stem cells) by a factor of about 48%, but a combination of the natural substances carnosine, blueberry extract, green tea extract and vitamin D3 was potent24. This combination increased the early stem cells by a factor of 68%24. It is gratifying to see how synergistic combinations of natural substances (with no significant adverse effects) can sometimes outperform expensive prescription drugs, such as GM-CSF, in this context.
The use of seaweed in stimulating stem cell production has been ascribed to the ability of complex carbohydrates (fucoidans), contained with seaweed, to promote adult stem cell release from bone marrow stores into the peripheral blood or general circulation36-39. While fucoidans may bind with fibroblast growth factors and promote angiogenesis (new blood vessel growth), they have immuno-modulating, anti-inflammatory properties36-39. Angiogenesis (promotion of the growth of new blood vessels) has been highlighted as an important process in human stem cell engraftment1.
Chris D. Meletis, ND has drawn attention to the stem cell enhancement potential of fucoidan found in marine algae36. This polysaccharide compound, found in abundance in Wakame seaweed, has been shown to exert influence on the mobilization of endothelial progenitor cells, associated with the incorporation of such cells into ischemic tissue. It would appear that fucoidan acts through the modulation of the activity of SDF-136-39. In addition, it appears that fucoidan has pro-angiogenic activity which is very important in the process of repair of tissue damage36-39. The benefits of fucoidan appear to be particularly important in general tissue repair processes and cardiovascular health36.
In brief, it appears that there is a group of miscellaneous botanicals that may have value in induction of ASC mobilization or recruitment. Examples of these botanical agents include: green tea polyphenols40, blueberries41, other anthocyanidins and vegetables including spirulina, spinach, grape seed extract and Ashwagandha25. There are many potential mechanisms, whereby these botanicals can exert benefit in IASCR. For example, green tea polyphenols have anti-aging, angiogenic, anti-cancer and cell protective benefits. Blueberries, spinach and strawberries contain compounds that modulate cell-signaling cascades42. They have regenerative effects on certain cell populations42. Other botanicals that have been mentioned will provide, at least, antioxidant and variable cell regulatory potential25.
While it is recognized that adult stem cells are often “tissue specific,” they may be somewhat limited by an ability to replace specific cell types that are damaged. However, adult stem cells retain a degree of pluripotency (multipotent) that permits them to differentiate (form a special adult cell identity) into somatic cell types that are present in different organ tissues. Clearly, further investigations of the ability of nutraceuticals to support adult stem cell proliferation and mobilization are required25.

Summarizing the Concepts of IASCR
Table 2 simplifies the concepts proposed for the use of IASCR (Holt, 2009).

Mobilize ASC from bone marrow and other niche locations
Increase circulation of ASC with semi-continuous, safe stimuli. (Herbs, botanicals and nutrients are preferred to drug approaches, (they cost less, and have less side effects)
Protect ASC from oxidative damage
Encourage homing to desired target organ (?)
In-vivo assistance in the differentiation of ASC to replace cell types of the diseased organ has to be developed (The “human petri-dish” approach).
A body of research demonstrates that human bone marrow ASC are able to “home in” on diseased organs and differentiate into many cell types.

Table 2. The concepts of how to utilize the Induction of Adult Stem Cell Recruitment (IASCR) (Holt, 2009)25.
There are several unresolved issues in the proposals that I make concerning IASCR (Table 2). While it seems clear that ASC release and migration occurs following the use of several nutraceuticals, the question of deployment of mobilized ASC to organs has not been evaluated. Diseased or ailing tissues are known to produce many chemo-attractants for circulating stem cells17-19. Intrinsic in the progress of IASCR following stem cell mobilization, is a plausible proposal that homing of progenitor or ASC will occur in a reproducible manner. Questions arise concerning the engagement and recruitment of ASC at target sites. This process must be followed ideally by desired proliferation and engraftment. These matters remain to be clarified.
Within the concepts of IASCR rests the possible applications of some substances that can assist in promoting the in-vitro differentiation of mobilized stem cells towards desired cell types, in order to regenerate specific disease organs. Many physical or chemical manipulations are applied in the laboratory to transform (manipulate) harvested stem cells, in order that they may be stimulated to differentiate along certain cell lineages. Perhaps these “petri-dish” manipulations of stem cells can be applied in an overall process of IASCR, making the circumstances somewhat like a “human petri-dish” of stem cell differentiation? These matters are quite speculative at present, but it is known that the simple application of key nutrients can have major effects on pathways of differentiation taken by cultured stem cells in-vitro1.
In order to use nutraceutical induction of ASC mobilization or recruitment (IASCR), one must propose a rational basis for this novel procedure25. The nutraceuticals that have been shown to have mobilization and protection potential for ASC have potential health benefits that extend beyond the act of IASCR25, including their antioxidant potential and cell regulatory potentials. While I acknowledge that much further research may be required to validate my proposals for the routine use of IASCR, cumulative evidence to date reinforces this promising novel approach for disease management and, perhaps, the promotion of longevity25.
Formulations for Stem Cell Support
In this overview, the power of synergy among nutrients, herbals and botanicals in the facilitation and mobilization of ASC and their potential recruitment by damaged or ailing tissues have become apparent. It seems prudent to conclude that the use of a single stem cell mobilizing factors alone, e.g. AFA, is not likely to be as effective as synergistic combinations of nutraceuticals. These circumstances make several proposals on single or limited combinations of nutrients with botanicals somewhat obsolete. Cumulative scientific studies appear to support the use of more complex synergistic nutraceutical formulations, with greater potential functionality. These proposals supersede several existing patents on stem cell mobilization. Table 3 proposes a complex nutraceutical formulation that can be used in the support of stem cells.

Fatty acids: linolenic and oleic acid
Antioxidant protectors including anthocyanidins (blueberry and beet root), spirulina, green tea polyphenols, OPC and fucoxanthin
Key nutrient support with Vitamin D3 for cell proliferation and nutritional support of bone marrow function with Vitamin B12 and folate.
Specific evidence-based stem cell releasers, e.g. AFA, blueberry, Vitamin D3 and fucoidan.

Table 3. One of several proposals for complex synergistic formulations to induce adult stem cell mobilization and antioxidant protection, with ancillary functionality (Courtesy of Holt MD Technologies,
More than a decade of research exists in the use of natural compounds to assist in the mobilization or recruitment of endogenous ASC. I propose the concept of IASCR with the use of nutritional support for stem cell function25. This approach appears feasible and readily applicable without any significant risks of adverse effects. This non-invasive area of stem cell technologies appears quite attractive, given the lack of portability and the limitations of current stem cell procedures that require a combination of advanced clinical skills and sophisticated laboratory support.


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