Chinese Journal of Tissue Engineering Research ›› 2013, Vol. 17 ›› Issue (49): 8512-8519.doi: 10.3969/j.issn.2095-4344.2013.49.008

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Salidroside and cholestatic serum induce differentiation of bone marrow mesenchymal stem cells into hepatocytes

Du Chao, Jiang Ming-de, Zeng Wei-zheng, Gao Yong   

  1. Department of Gastroenterology, General Hospital of Chengdu Military Command, Chengdu  610083, Sichuan Province, China
  • Revised:2013-09-02 Online:2013-12-03 Published:2013-12-03
  • Contact: Jiang Ming-de, Master, Chief physician, Professor, Department of Gastroenterology, General Hospital of Chengdu Military Command, Chengdu 610083, Sichuan Province, China jiangmd88@yahoo.com.cn
  • About author:Du Chao★, Master, Attending physician, Department of Gastroenterology, General Hospital of Chengdu Military Command, Chengdu 610083, Sichuan Province, China 176147067@qq.com

Abstract:

BACKGROUND: A large number of experiments have confirmed that bone marrow mesenchymal stem cells can differentiate into hepatocytes under the induction of cytokines and specific micro-environment, and have been widely used in clinical alternative treatment for terminal liver disease, but the optimal inducing conditions are unclear.
OBJECTIVE: To explore the possibility and validity of differentiation of rat bone marrow mesenchymal stem cells into hepatocytes with a culture system containing salidroside and cholestatic rat serum in vitro.
METHODS: Bone marrow mesenchymal stem cells were isolated by plastic adherence from the whole bone marrow of health rats, and cell phenotypes were identified using the flow assay; cholestatic serum was prepared by common bile duct ligation. Passage 3 bone marrow mesenchymal stem cells were randomly divided into three groups for in vitro induction by the different culture systems: blank control group: basic medium plus 5% cholestatic serum; salidroside group: basic medium plus 5% cholestatic serum plus 30 µmol/L salidroside; positive control group: basic medium plus 5% cholestatic serum plus 20 µg/L hepatocyte growth factor. Changes of cell morphology during culture time were observed in each group, reverse transcription-PCR assay and western blot assay were used to expression of hepatocyte-specific proteins.
RESULTS AND CONCLUSION: The bone marrow mesenchymal stem cells highly expressed CD90, CD105, but did not express CD45, CD14, CD34, and CD79a. Polygonal and binucleate cells appeared in the three groups during the procedure of induction. The mRNA and protein expression of alpha-fetoprotein and albumin emerged in the three groups on the 7th day; in the same period, the lowest expression ratio was in the blank control group (P < 0.05), while there was no significant difference between the salidroside and positive control groups (P > 0.05). Combination of salidroside and cholestatic serum can effectively induce bone marrow mesenchymal stem cells differentiating into hepatocytes.

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

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