Optimizing Neuronal Differentiation From Human Umbilical Cord Derived Wharton Jelly-Mesenchymal Stem Cells

Sharma, Avinash (2015) Optimizing Neuronal Differentiation From Human Umbilical Cord Derived Wharton Jelly-Mesenchymal Stem Cells. Masters thesis, Indian Institute of Science Education and Research Kolkata.

PDF (MS dissertation of Avinash Sharma) Avinash_Sharma_13IP043.pdf - Submitted Version Restricted to Repository staff only Download (1MB)

Abstract

Since past few decades Mesenchymal Stem Cells (MSCs) have evolved as a new field of research in the areas of regenerative medicine. Particularly, their lack of immunogenicity, tumorigenicity and ability to perform immunomodulatory as well as anti-inflammatory functions make MSCs a potential source of regenerative and cell-based transplantation therapy. Researchers have found that MSCs derived from various sources have ability to differentiate into multiple types of cells like neurons, cardiomyocytes, hepatocytes, osteoblasts, insulin producing cells etc. A few findings have showed that MSCs specially, Wharton-Jelly derived MSCs (WJ-MSCs) have propensity to differentiate into specific neuronal lineage of cells like dopaminergic neurons, oligodendrocytes, astrocyte like cells which can be really helpful in various neurodegenerative disorders. However, although MSCs are multipotent in nature and have been found to differentiate into multiple types of cells using different kind of chemically and biologically active compounds, the underlying mechanism and signaling is not well understood. We have treated WJ-MSCs with basic fibroblast growth factor (bFGF) and forskolin as induction factors for 14 days. bFGF and forskolin both are thought to induce neural fate of MSCs as bFGF generates neural precursor cells and forskolin is a chemical compound which triggers cAMP signaling pathway by acting on adenylyl cyclase and increasing intracellular cAMP level followed by Protein Kinase A (PKA) activation. This all enhances neural differentiation of MSCs. Using bFGF+forskolin we have found morphological changes in WJ-MSCs where they showed typical dendritic morphology similar to neurons. PCR data of induced WJ-MSCs showed the expression of mature neuronal markers like NF-M (Neurofilament-M), Neuron Specific Enolase (NSE) and also Nurr1 (Nuclear Receptor related 1 protein) which is known to play a key role in dopaminergic system maintenance. However, we did not detect any expression of Map2. Also, early progenitor markers such as Sox1, Pax6 and Nestin, were not expressed in our induced cells. Preliminary data using Neurobasal/B27 supplement media and bFGF has shown neurosphere like formation by WJ-MSCs. Our future aim is to improve the differentiation efficiency of WJ-MSCs into neurons under different in vitro conditions, e.g.hypoxia. We also want to study the underlying signaling mechanism behind differentiation.

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