Intracellular Ca2+is involved in survival, proliferation and differentiation of bone marrow-derived mesenchymal stem cells into hepatocytes
10.3969/j.issn.2095-4344.2013.40.002
- VernacularTitle:Ca2+参与骨髓间充质干细胞存活增殖及向肝细胞的诱导分化
- Author:
Shuxian JIAO
;
Bin HU
;
Lin ZHAO
;
Xiaohua LIU
;
Zhihui FENG
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2013;(40):7028-7033
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:The mechanism of differentiation and proliferation of bone marrow-derived mesenchymal stem cells remains unclear. In addition, issues such as how signal pathways such as Ca2+and bone marrow-derived mesenchymal stem cellproliferation and differentiation signals form complex signal network remain poorly understood.
OBJECTIVE:To investigate the effect of Ca2+in the induced differentiation of bone marrow-derived mesenchymal stem cells into hepatocytes.
METHODS:Bone marrow-derived mesenchymal stem cells were isolated from rat bone marrow using whone bone marrow adherence method, purified, amplified, and induced with hepatocyte growth factor. [Ca2+]i in the directional differentiated bone marrow-derived mesenchymal stem cells and control bone marrow-derived mesenchymal stem cells were detected with flow cytometry. Bone marrow-derived mesenchymal stem cells induced with hepatocyte growth factor were mixed with nimodipine of different concentration, and cells were divided into three groups:hepatocyte growth factor+nimodipine 10 mg/L, 50 or 100 mg/L groups. cellgrowth was observed with inverted phase contrast microscope and alpha 1-antitrypsin expression of the cells was confirmed by immunocytochemistry. The calcineurin M and the activation of extracellular signal regulated kinase pathway was detected by reverse transcription-PCR and western blotting, respectively.
RESULTS AND CONCLUSION:[Ca2+]i in the directional differentiated bone marrow-derived mesenchymal stem cells was higher than in the control group (P<0.05). After addition of a larger dose of nimodipine, no differentiation of cells was obeserved and growth of bone marrow-derived mesenchymal stem cells was getting worse. There were few alpha 1-antitrypsin positive cells in the nimodipine groups. Calcineurin Mexpression was significantly increased in directional differentiated bone marrow-derived mesenchymal stem cells and smal dose of nimodipine than the controls (P<0.05). However, no significant difference was found among middle, high dose nimodipine and control groups (P>0.05). These findings indicate that Ca2+could participate in the differentiation of bone marrow-derived mesenchymal stem cells into hepatocytes incuded with cytokines, and also maintain the survival and proliferation of bone marrow-derived mesenchymal stem cells.
