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What to know before getting stem cells

Dr. John Hughes, DO

Aspen Integrative Medicine

July 6th, 2017

Disclaimer:
I have no material interest or investment in any stem cell companies or products.

  • I. Introduction to stem cells
  • II. Mesenchymal stem cells
  • III. Pluripotent stem cells
  • IV. Making a stem cell decision

Embryonic Stem Cells

  • Derived from the fetus
  • Requires special regulatory approval
  • Mostly used for research purposes
  • Not readily available
  • Expensive
  • Not autologous

Adult Stem Cells

  • Derived from bone, adipose, or blood
  • Requires physician expertise and quality control
  • Mostly used for regenerative and cosmetic purposes
  • Readily available
  • Less expensive
  • Autologous use is permitted in US

Totipotent Stem Cells

  • Derived from the embryo
  • Forms all cell types in the body
  • Used for early embryonic development
  • “Baby cells” (from a zygote) that becomes a pluripotent cell

Pluripotent Stem Cells

  • Derived from the embryo or blood
  • Forms all cell types in the body except the embryo or the placenta
  • Does not have a specialized trajectory of development
  • “Youthful cell” with great ability to differentiate into other cell types

Multipotent Stem Cells

  • Derived from cord blood, blood, bone marrow, fat and muscle
  • Forms only cell types from the mesoderm
  • Has a development trajectory towards a specific type of cell
  • “Teenage cell” already differentiated into it’s target adult cell type

Mesenchymal Stem Cells (MSCs)

Discovery

  • Alexander Friedenstein discovered mesenchymal stem cells in mice (Mus musculus).
  • From 1966 through 1987, Friedenstein provided evidence that stem cells from bone marrow can differentiate into mesenchymal tissues
  • Since then, the cell potency of mesenchymal stem cells differentiation has been a cause of debate
  • Are they truly multipotent or unipotent?

https://embryo.asu.edu/pages/mesenchyme

Defined

  • Mesenchyme: loose cells embedded in the extracellular matrix
  • a mesh of proteins and fluid that allows cells to migrate easily
  • Directs development of morphological structures during the embryonic and fetal stages
  • connective tissues, bones, cartilage, lymphatic and circulatory systems
  • Carry over 480 growth factors and are attracted to target tissues of inflammation
  • Primarily isolated from fat or bone marrow through a time-insensitive, invasive process
  • Human fat (adipose tissue) has about 10x more stem cells than bone marrow

https://embryo.asu.edu/pages/mesenchyme

How They Work

  • Derived from pluripotent stem cells, have already partially differentiated, and they continue specializing as they develop
  • Must be activated appropriately – often mixed with human plasma
  • Considered multipotent because their specialization potential is limited to one or more cell lines
  • Current research suggests multipotent cells are able to go beyond the boundaries of producing one specific cell type but do so infrequently and only under narrow conditions

http://www.explorestemcells.co.uk/multipotentstemcells.html

  • Modulate endogenous tissue and immune cells
  • Actively interact with nearby cells
  • Observed benefits of MSC therapy may result from the relinquishment of their molecular contents upon administration
  • Therapeutic effects are short-lived
  • “Recent studies have suggested that less than 1% of systemically administered MSCs persist for longer than a week following injection.”
  • Limited in numbers – unlikely that true MSCs circulate peripherally (~0.01% of mononuclear bone marrow cells)

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3759519/

Clinical Indications

  • Allogenic bone marrow transplants (from the same species to another of the same species) clinically used since the 1980s
  • Autologous fat and bone marrow transplantations (from the same individual back to same individual) can be used to support target tissues that are not from the same cell type
  • E.g., injecting fat MSCs into joints orthopedically to support the growth of new cartilage
  • These MSC’s do not develop into new cartilage cells – they provide growth factors, reduction in inflammation, and immune modulation that may support joint health
  • They are already on a development trajectory and their effects on unique target tissues are mostly paracrine (effecting nearby cells)
  • Outside the US, MSCs can be cultured for several weeks to build up the cell counts
  • The idea is that with more MSCs, more target tissues will benefit
  • Challenge: they grow older the more times that they replicate so they are less effective
  • MSCs are generally best used for transplantation into similar tissues from which they derive
  • E.g., MSCs from fat are best transplanted into areas in need of fat replacement
  • (breast augmentation, subcutaneous fat areas of the body – facial, lip, buttocks transplants)
  • Most effective clinical use of MSCs:
  • Same tissue transplantation (bone marrow to bone marrow, fat to fat)
  • Joint conditions (if related to an autoimmune or systemic inflammation)
  • Autoimmune disorders and systemic inflammatory conditions (see table on next slide)

Dangers and Side Effects

  • Harvesting of bone marrow and fat MSCs is unpleasant for the patient
  • There is a limited number of times one can extract and use fat MSCs
  • Many patients have to repeat the procedure to gain significant benefit
  • MSCs reduce inflammation for a time period of 6 months to 2 years but have limited regenerative benefits
  • Are generally designed to affect one germ layer and tissues derived from that mesodermal layer – and under most conditions are unipotent (have the capacity to differentiate into only one cell type)
  • Because of the immunomodulatory effects of these MSCs, they predispose the patients more infections or even cancer
  • After MSC infusions were used to treat nine patients suffering from GvHD, three developed viral infections
  • Immunosuppression by the MSCs had caused a reduction of immuno-surveillance to viruses

https://www.ncbi.nlm.nih.gov/pubmed/16604097

  • MSCs, when administered in rats, can engraft in the renal tubules and mal-differentiate into adipocytes that hinder normal function of the kidney and lead to chronic kidney disease

https://www.ncbi.nlm.nih.gov/pubmed/17460140

Pluripotent Stem Cells

Discovery

  • Henry Young et al. (2004) demonstrated connective tissue (including blood) contains reserve precursor cells
  • Reserve precursor cells consist of: tissue-specific progenitor cells, germ-layer lineage stem cells, and pluripotent stem cells
  • Tissue-specific progenitor cells can be unipotent or multipotent
  • Progenitor cells can only double 50–70 times while germ-layer lineage stem cells and pluripotent stem cells have a much greater lifespan
  • Pluripotent stem cells were thought only to exist in embryonic stem cells until Dr. Young’s discovery of them in the peripheral blood in the late 20th century

Defined

  • Precursor cells can be
  • Tissue-specific progenitor cells
  • Lineage-committed (ectodermal, mesodermal, and endodermal) germ-layer lineage stem cells
  • Lineage-uncommitted pluripotent epiblastic-like stem cells
  • What we are interested in today are the lineage uncommitted pluripotent stem cells  (some researchers call these cells, blastomere-like stem cells)

https://mail.google.com/mail/u/0/#search/henry+young+/13b421977b3e5c33?projector=1

How They Work

  • Understanding lineage uncommitted pluripotent stem cells requires an understanding of the germ layers
  • Adult pluripotent stem cells can be induced to form cells from the three primary germ-layer lineages (i.e., ectoderm, mesoderm, and endoderm).
  • Results from neuronal, hematopoietic, diabetic, chondrogenic, osteogenic, myogenic, and cardiogenic studies demonstrate that adult pluripotent stem cells can be induced to undergo directed lineage induction.
  • The activation of quiescent precursor cells is a potential component of tissue restoration.
  • Quiescent stem cells also assist the tissue-committed progenitor cells in forming the missing tissues
  • Originate in bone marrow and present in peripheral blood
  • Contain a unique marker that can be used to select them for both diagnostic and therapeutic procedures
  • In abundance in peripheral blood and in reproductive tissue secretions

Clinical Indications

  • Lineage uncommitted pluripotent stem cells can be used to form any tissues in the endoderm, mesoderm, or ectoderm
  • Treatment of a wide range of degenerative diseases in both humans and animals including, but not limited to:
  • Diabetes, osteoarthritis, osteoporosis and Alzheimer’s disease, to name a few, as well as regenerative applications associated with aging
  • TBI studies: When used in conjunction with hyperbaric oxygen therapy, intranasal and IV pluripotent stem cells (derived from blood plasma), after activation, have been shown, in case studies, to positively address post-concussive symptoms secondary to TBI: memory, sleep, mental fatigue, mental clarity, libido, motor function and balance.
  • Could be shown useful in replacing bone marrow in post-cancer treatment

Making a stem cell decision

  • Bone Marrow
    • Cost: $3,000 – $10,000
    • Recovery time: One Month
  • Adipose (Fat)
    • Cost: $6,000 – $15,000
    • Recovery time: One Month
  • Blood Based
    • Cost: $3,500 – $4,000
    • Recovery time: Less than a week

References

Cell Applications. https://www.cellapplications.com/stem-0

Cellular Differentiation. http://oerpub.github.io/epubjs-demo-book/content/m46036.xhtml

Kunter, U., Rong, S., Boor, P., Eitner, F., Müller-Newen, G., Djuric, Z., … & Milovanceva-Popovska, M. (2007). Mesenchymal stem cells prevent progressive experimental renal failure but maldifferentiate into glomerular adipocytes. Journal of the American Society of Nephrology18(6), 1754-1764.

MacCord, K. (2012). Mesenchyme. The Embryo Project Encyclopedia. Retrieved from https://embryo.asu.edu/pages/mesenchyme

Mesenchymal Stem Cell Reagents. Retrieved from http://www.sigmaaldrich.com/life-science/cell-biology/cell-biology-products.html?TablePage=22692887

Murnaghan, I. (2016). Multipotent stem cells. Explore Stem Cells. Retrieved from http://www.explorestemcells.co.uk/multipotentstemcells.html

Parekkadan B, Milwid JM. Mesenchymal Stem Cells as Therapeutics. Annual review of biomedical engineering. 2010;12:87-117. doi:10.1146/annurev-bioeng-070909-105309.

Stout, C. L., Ashley, D. W., Morgan, J. H., Long, G. F., Collins, J. A., Limnios, J. I., … & Young, H. E. (2007). Primitive stem cells residing in the skeletal muscle of adult pigs are mobilized into the peripheral blood after trauma. The American Surgeon73(11), 1106-1110.

Sundin, M., Örvell, C., Rasmusson, I., Sundberg, B., Ringden, O., & Le Blanc, K. (2006). Mesenchymal stem cells are susceptible to human herpesviruses, but viral DNA cannot be detected in the healthy seropositive individual. Bone marrow transplantation37(11), 1051-1059.

Tithon Biotech (n.d.). Minutevideo retrieved from http://mv.pac.io/post/55b2e2b41e1818650cc7b872

Tithon Human Sciences (2015). Peripheral blood derived pluripotent stem cell technology. Rhttp://aspenintegrativemedicine.com/wp-content/uploads/Tithon-Human-Sciences-Ortho-Case-Study.pdf

Young, H. E., Duplaa, C., Romero-Ramos, M., Chesselet, M. F., Vourc’h, P., Yost, M. J., … & Tamura-Ninomiya, S. (2004). Adult reserve stem cells and their potential for tissue engineering. Cell biochemistry and biophysics40(1), 1-80.

Young, H. E., & Black, A. C. (2004). Adult stem cells. The Anatomical Record Part A: Discoveries in Molecular, Cellular, and Evolutionary Biology276(1), 75-102.