Increasing the number and survival of hESC and the derived lineages in tissue culture.
Our research for methods to protect the cells during aging revealed that N-t-butyl hydroxylamine (NtBHA, active form of a drug in clinical trial for stroke) and methylene blue (MB, a well known drug) significantly extend the vigor and lifetime of cells. MB or NtBHA at very low concentrations delay senescence of the cells; increase the number of the cells by 6-7 million cells/week; enhance cellular resistance to oxidative stress and toxic agents; and improve energy production. When fed to old rats MB and NtBHA enhance cognitive function and improve muscle strength. Preliminary observations suggest that NtBHA and MB, accomplish this, in part, by improving the quality of the mitochodnria, which are the cellular source for energy. MB and NtBHA also lowered the levels of oxidative damage to the cell compnents.
We will use MB and NtBHA to improve the maintenance of hESC and the derived neuronal lineages in culture. We are expecting MB and NtBHA to improve the culturing of hESC and derived cells when added to the established growth media. Thus, we hypothesize that maintaining hESC and derived neuronal lineages with MB or NtBHA considerably increases the total number of cells, efficiency of derivation, and survival in the tissue culture conditions.
If the goals of this proposal are successfully accomplished, then the quality and the number of the available stem cell units for cell therapy should improve. Additionally, the storage of stem cells at –80 °C will be made safer.
A future longer-term objective of our proposed research is to test whether MB and NtBHA promote the function of endogenous stem cells when administered in experimental animal models. The basis for this hypothesis is that MB and NtBHA enhanced the cognitive function and improved muscle strength in rats. A one possible explanation for this effect is that MB and NtBHA, in part, improve the function of endogenous stem cells in the rat’s tissues (e.g. increased the secretion of specific hormones). Additionally, MB and NtBHA may enhance the metabolic activity of the somatic cells adjacent to the stem cells, which, based on recent research, should preserve the function of the endogenous stem cells in the rat’s tissues. Promoting the function of endogenous stem cells has been proposed as a therapeutic strategy to prevent and cure several diseases in human. A pharmacological approach to enhancing the function of native stem cells in tissue (e.g. the use of MB and NtBHA) to prevent disease (e.g. Alzheimer disease) may complement stem-cell transplantation therapy.
In order for the objectives of Proposition 71 to be accomplished and patients in Californian to benefit from the use of human embryonic stem cells (hESC) to regenerate tissues to replace tissues damaged by disease and injury, researchers must first be able to grow and maintain sufficient numbers of healthy and pure hESC and their derived lineages. Current hESC research is impeded by the technical difficulties and safety concerns in getting hESC, especially the derived lineages, to grow in adequate numbers under laboratory conditions and being able to maintain the cells in a healthy state. We are proposing to test the effect of two compounds, which have been shown to improve the health of aging cells from old rats, without harmful side effects. These two compounds appear to accomplish this by preventing the dysfunction of key organelles known as mitochondria, which produce energy that cells use for all biological processes, including cell division, cell repair, and the production of enzymes and hormones. If our research is successful, it will help speed up basic hESC research, use of hESC in the private biotechnology sector, and cell replacement therapy in California. This research, in conjunction with the ongoing research at many of California’s institutes, should help position California at the forefront of stem cells research.