Proliferation Capacity and Expression of Cell Cycle Genes in Normal and Gestational Diabetes Affected WJMSCs
Proliferation Capacity and Expression of Cell Cycle Genes in Normal and Gestational Diabetes Affected WJMSCs
Nabila Rasheed1, Shumaila Usman2*, Madeeha Sadiq3, Bushra Wasim3, Kanwal Haneef4, Hafsa Shafqat3, Tabinda Urooj3 and Saman Rashid4
ABSTRACT
Wharton’s jelly derived mesenchymal stem cells (WJMSCs) are a well-known source for the regenerative approach. Current investigation shows that the metabolic disorders like gestational diabetes mellitus (GDM) are not only responsible for several disease factors in adult life but also have adverse effects on perinatal environment as well as on the cellular properties of WJMSCs. This study was designed to determine the effects of gestational diabetes mellitus (GDM) on proliferation of WJMSCs (GDM-WJMSCs). Mesenchymal stem cells were isolated from Wharton’s jelly tissue of normal(healthy) samples and from the GDM affected sample. The isolated cells were characterized for the presence of stem cells markers by Immunocytochemistry. After characterization, the analysis of GDM effect on WJMSCS proliferation rate was analyzed by calculating the population doubling time of the cells at different passages (P0-P3). Moreover, the expression of the important cell cycle genes (CDCA2, CDCA8, CDC20, and CCNA2) was analyzed by quantitative PCR to determine the effect of GDM at gene level. The isolated cells expressed the stem cell specific marker CD90 which confirmed that the isolated cells are WJMSCs. Population doubling time (PDT) was found to be increased in the early passages (P0-P1) in GDM-WJMSCs as compared to control WJMSCs. However, at later passages (P2, P3 stages) PDT was decreased and an improvement in the proliferation rate was observed. The fold change expression of cell cycle genes was found to be decreased in GDM-WJMSCs as compared to control WJMSCs. This study concluded that the GDM affect the expression of cell cycle genes, hence the cells proliferate slow as compared to the normal cells. The GDM affected cells had a high population doubling time in the early passages but when the cells were grown in vitro in controlled environment the cells proliferate efficiently, suggesting that the GDM affected WJMSCs can also be utilized for the regenerative purpose after improving the proliferation rate of the cells.
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