Chinese Journal of Tissue Engineering Research ›› 2013, Vol. 17 ›› Issue (40): 7028-7033.doi: 10.3969/j.issn.2095-4344.2013.40.002

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Intracellular Ca 2+ is involved in survival, proliferation and differentiation of bone marrow-derived mesenchymal stem cells into hepatocytes 

Jiao Shu-xian, Hu Bin, Zhao Lin, Liu Xiao-hua, Feng Zhi-hui   

  1. Institute of Transfusion Medicine, Qingdao Blood Center, Qingdao  266071, Shandong Province, China
  • Online:2013-10-01 Published:2013-10-31
  • Contact: Feng Zhi-hui, Master, Attending physician, Institute of Transfusion Medicine, Qingdao Blood Center, Qingdao 266071, Shandong Province, China zhaolinqd@163.com
  • About author:Jiao Shu-xian, Associate chief physician, Institute of Transfusion Medicine, Qingdao Blood Center, Qingdao 266071, Shandong Province, China qd_hla@163.com
  • Supported by:

    the Qingdao Science and Technology Support Plan Project, No. 09-1-1-25-nsh*

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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 Ca 2+and bone marrow-derived mesenchymal stem cell proliferation and differentiation signals form complex signal network remain poorly understood.
OBJECTIVE: To investigate the effect of Ca 2+ 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. [Ca 2+ ]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. Cell growth 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: [Ca 2+ ]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 M expression was significantly increased in directional differentiated bone marrow-derived mesenchymal stem cells and small 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 Ca 2+ 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.

Key words: stem cells, mesenchymal stem cells, hepatocyte growth factor, cell differentiation

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