Neil Riordan PhD on stem cell expansion in stem cell therapy

Stem Cell Pioneers featured Dr. Riordan in its February installment of “Ask the Doctor”, a monthly segment that features stem cell scientists and doctors answering questions from readers about stem cell therapy.

Over the next several days, we will share these questions and Dr. Riordan’s answers with our readers.

Question: Are there some conditions such as neurological ones that respond better when the cells are greatly expanded? Is a high quantity essential for success or is that something that may be more of a selling point at some clinics? I have also seen this advertised for COPD and other conditions. It’s almost like the more cells the better, but I would like your opinion.

Dr. Riordan’s Answer: That really depends on the quality of the cells after expansion. If they are still robust, not senescent, and still have a good secretion profile, then the more the better may be useful up to a point. If you take a small pool of starter cells and expand them to exhaustion, then I don’t think you are going to have a very good product. The MSCs used in Panama are not expanded beyond passage 5—a point at which there is no senescence in the population and they have a robust cytokine secretion profile. In order to use only cells that meet our release criteria, cells from approximately one (1.2 to be exact) out of 10 donated umbilical cords are used.

Contrast that to cells from a patient’s own fat tissue that are expanded. Firstly, the starting cells may, and many times are not very robust—they secrete little or no beneficial cytokines or chemokines, and must be expanded to hilt in order to hit the cell number. Please see my answer to number 7 for more on this subject.

This brings up a slightly different, yet related topic. There has been a lot of talk at recent meetings about more defined endpoints for the cells being used, and I couldn’t agree more. There are MSCs from bone marrow, menstrual blood, fat tissue, umbilical cord (even different parts of the umbilical cord—around the blood vessels, from the Wharton’s jelly, from the subepithelium, from the cord blood itself—which are most likely contaminants from a bruised placenta rather than the blood), teeth, amniotic membrane, amniotic fluid just to name sources in the “we didn’t mess with mother nature” adult stem cell world. Add to that the infinite variables when you consider the age and physical condition of the donor, particularly when using adipose or bone marrow as a source material and we, as a field, could be saying almost anything by using the term, “mesenchymal stem cell.” I think it is time that there is standardization in the field beyond the current definition of expressing/not expressing certain surface markers and the ability to differentiate into fat, bone, and cartilage. That standardization could come from using endpoints such as “remaining proliferative capacity (the number of doublings achievable in culture from the treatment cell bank), the secretome, even if there is standardization of one or two molecules, such as HGF, or one of the prostaglandins.

In the future I believe the field will take it a step further by measuring, even by a surrogate marker, the potential effects of the cells on the target condition. In the case of autoimmunity the cells and their secretions could be tested for their capacity to modulate the immune system. In the case of inflammatory conditions, the cells and their secretions could be tested for the ability to control or block inflammation.

The Senescence-Related Mitochondrial/Oxidative Stress Pathway is Repressed in Human Induced Pluripotent Stem Cells

Embryonic stem cells possess the ability to propagate in
tissue culture indefinitely.  This is different than differentiated cells, for
example, skin cells which can only multiple in tissue culture approximately 50
times before undergoing senescence.  The ability of embryonic stem cells to
escape senescence is related to expression of the protein telomerase.  Usually
when cells multiply the ends of the chromosomes, called telomeres, progressively
reduce in size.  When the telomeres become critically short, the gene p53 is
activated, which is involved in instructing the cells to stop multiplying and
exist in a semi-alive state called senescence.  Tumor cells and embryonic stem
cells escape senescence by expressing the enzyme telomerase.  This enzyme
essentially allows cells to repair their telomeres by progressively adding new
nucleic acids.  Although much is known about senescence or lack thereof in adult
cells and embryonic stem cells, little research has been performed in whether
inducible pluripotent stem cells (iPS) can also escape proliferative
senescence.  In a recent publication this question was examined.

In a similar manner to embryonic stem cells, iPS cells were
shown to express high levels of the enzyme telomerase, and propagation in tissue
culture was achieved up to 200 passages without senescence occurring. 
Furthermore the investigators studied the mitochondrial stress pathway.  It was
found that somatic mitochondria within human iPSCs revert to an immature
ESC-like state with respect to organelle morphology and distribution, expression
of nuclear factors involved in mitochondrial biogenesis, content of
mitochondrial DNA, intracellular ATP level, oxidative damage, and lactate
generation. When iPS cells were differentiated into adult cells, mitochondria
within iPSCs demonstrated maturation and anaerobic-to-aerobic metabolic
modifications. This same finding was observed in embryonic stem cells. 

These data suggest that iPS cells possess several important
properties similar to embryonic stem cells, which further supports the
possibility of interchangeably using ES and iPS cells for experimental purposes.
The next question is whether iPS cells may be generated in large quantities so
that their mitochondria may be transferred to aged cells. 

Another interesting finding in the current study is that
the metabolic pathway used by both iPS and embryonic stem cells is analogous to
that found in cancer cells.  Therefore it will be interesting to follow studies
using iPS as a model of cancer.