Salidroside and cholestatic serum induce differentiation of bone marrow mesenchymal stem cells into hepatocytes / 中国组织工程研究

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 cellphenotypes 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 systemsblank control groupbasic medium plus 5%cholestatic serum;salidroside groupbasic medium plus 5%cholestatic serum plus 30 μmol/L salidroside;positive control groupbasic medium plus 5%cholestatic serum plus 20 μg/L hepatocyte growth factor. Changes of cellmorphology 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 (P0.05). Combination of salidroside and cholestatic serum can effectively induce bone marrow mesenchymal stem cells differentiating into hepatocytes.