| Summary: | 碩士 === 慈濟大學 === 生理暨解剖醫學碩士班 === 100 === Stem cells have certain features such as self-renewal and differentiation potential. Adult stem cells are minor populations found in adult organs, they cannot give rise to an organism and only differentiate to specific cell lineages, mesenchymal stem cells (MSC) belong to this group. MSC can differentiate into several lineages, including osteocytes, chondrocytes and adipocytes. Recently, unexpected plasticity has been attributed to mesenchymal stem cells, as they have been demonstrated to differentiate into a number of non-mesoderm-type cells such as cardiomyocytes and neuron cells. In this study, we induce rat MSC (rMSC) to differentiate into neuron-like cells and adipocytes to investigate the correlation between calcium and cell differentiation.
Calcium, a ubiquitous second messenger, regulates many cellular functions, including cell growth, differentiation, and apoptosis. Intracellular calcium homeostasis is maintained via calcium channels, calcium pumps and calcium exchangers. Store-operated calcium entry (SOCE) are the major source of intracellar calcium in non-excitable cells. Therefore, the aim of this study is to investigate the role of SOC channels in rMSC differentiation.
rMSC is induced to differentiate into adipocytes and neuron-like cells. We measure the difference of concentration between Control and differentiation groups using microspectrofluorometry. We find that calcium influx through SOC channels and calcium depletion is significantly increase in differentiated rMSC. Therefore, we use Western blot and Q-PCR to determene that whether the expressions of STIM-1, Orai-1 and TRPC1 is differences between Control and differentiation groups. Our finding show that the levels of STIM-1 expression is significantly increase, but the levels of Orai-1 and TRPC1 expression have no significant difference between Control and differentiation groups. Inhibition of SOCE by a treatment with 2APB can blocked rMSC differentiation. Next, we measured the expression of IP3R1, IP3R2 and IP3R3 and find a significantly increase in IP3R2 after differentiation. Our results suggest that STIM-1 may play a critical role in the differentiation process.
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