Modulation of multiple differentiation potential in mesenchymal stem cells via an intercellular calcium signaling pathway

[Speaker] Minae Takahashi:1
[Co-author] Shuang Liu:1, Erika Takemasa:1, Masaki Mogi:1
1:Pharmacology, Ehime University School of Medicine, Japan

Background: Multipotent mesenchymal stem cells (MSCs) are widely used in regenerative medicine to repair damaged tissues. Regulating calcium release-activated calcium (CRAC) channel-mediated intercellular calcium signaling is critical to the functional modulation of MSCs. CRACM1 in the plasma membrane is an important molecular regulator of the CRAC channel. Our study aimed to elucidate the possible effects of CRAC channel regulation on the multiple differentiation potential of MSCs.
Methods: To increase calcium influx, we overexpressed CRACM1 by transfecting PCDNA3-CRACM1 into MSCs (m1-MSCs). An increased CRACM1 mRNA level was confirmed by RT-PCR and calcium influx was monitored using calcium imaging. YM-58483 was used as a CRAC blocker. The inhibitory effect of YM-58483 was verified by imaging. Wild-type MSCs, m1-MSCs, and YM-58483-treated MSCs were differentiated to adipocytes or osteocytes. Fatty acid-binding protein 4 was used as an adipocyte biomarker and osteocalcin was used for the detection of osteocytes.
Results: YM-58483 inhibited calcium influx in MSCs in a dose-dependent manner, as demonstrated by calcium imaging of fura-4-loaded cells. YM-58483 suppressed the differentiation of osteocytes, but not of adipocytes. In contrast, overexpressing CRACM1 had no effect on the differentiation of osteocytes, but suppressed adipogenic differentiation compared with that observed in wild-type cells.
Conclusions: The results suggest that MSC differentiation potential is related to intercellular calcium signaling pathways. In the future, we aim to research chondrocyte differentiation and, eventually, to carry out experiments in animal models to further contribute to the growing field of regenerative medicine.

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