Characterizing the role of Non Muscle Myosin II in proliferation and the Effect of Serum Deprivation on Maintenance of Pluripotency of Human Wharton’s Jelly Mesenchymal Stem Cells

Sharma, Tulika (2013) Characterizing the role of Non Muscle Myosin II in proliferation and the Effect of Serum Deprivation on Maintenance of Pluripotency of Human Wharton’s Jelly Mesenchymal Stem Cells. Masters thesis, Indian Institute of Science Education and Research Kolkata.

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Abstract

Wharton’s Jelly Mesenchymal Stem Cells (WJMSC) holds the potency of becoming a tool for cell-based therapy. Their multipotent, immunomodulatory, homing capacity, non-tumorogenic character and easy availability give these cells an edge over other forms of stem cells i.e. embryonic stem cells and adult stem cells. Stem cells, in general, are present in low number in their niche. To be able to use WJMSCs for clinical purpose, we need to expand the culture in vitro. Non muscle myosin, being a motor protein involved in cell division (cytokinesis), cell migration, and cell-cell and cell-matrix adhesion, becomes a contending candidate for investigating its role in proliferation capability of WJMSCs. blebbistatin, an inhibitor of NMII activity, inhibits the ATPase activity by interfering with the phosphate release process which reduce the affinity of NMII to bind with actin filament. In this study, we have isolated the WJMSCs from human umbilical cord and characterized them through immunophenotyping and differentiating the cells to adipocyte and osteocyte lineages. Through this study, it has been shown that disruption of actomyosin cytoskeleton leads to reduced proliferation of WJMSCs. WJMSCs in presence of blebbistatin exhibit a profound change in cell morphology i.e. a loss in spindle shape, flattened out cell shape, formation of stress fibre. Flow cytometry data and senescence data confirm that the WJMSCs grown in presence of blebbistatin show a G0/G1 growth arrest with cells going to premature senescence. Serum deprivation at the site of infection/inflammation pose a major issue towards the success of cell based therapies. in vitro studies in bone marrow MSC (BMMSC) has shown that when BMMSC were subjected to serum deprived condition, it led to the selection of a population of cell with higher expression of pluripotent markers. Hence we wanted to study whether WJMSC are able to survive in serum deprived condition and if yes then how do they behave in terms of stem cell marker expression. Our study has shown that when WJMSC were subjected to serum deprived condition they also led to a selection of population that expressed higher levels of pluripotent markers.

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