Rat bone marrow masenchymal stem cells differentiate into neuron-like cells and glial-like cells under hippocampal neuron conditioned medium in vitro Compared with basic fibroblast growth factor medium and serum-free medium / 中国组织工程研究

ABSTRACT

BACKGROUND: There are few reports addressing the differentiation of bone marrow mesenchymal stem cells (BMSCs) into neurons, and the uncertainties mainly focused on the differentiated neurons had neuron morphology, but did not have neuron function. OBJECTIVE: To investigate the feasibility of rat bone marrow mesenchyma stem cells (BMSCs) differentiation into neuron-like cells and glial-like cells under rat hippocampal neuron's conditional medium. METHODS: Rat BMSCs at passage 5 were divided into 4 groups. The medium of hippocampal neurons and glial cells was added in the conditioned medium group. The Dulbecco's modified Eagle's medium containing bFGF was added in the basic fibroblast growth factor (bFGF) group. The serum-free medium containing Neurobasal and B27 was added in the serum-free medium group. The DMEM supplemented with fetal bovine serum was added in the negative control group. 12 and 24 hours following induction, neuron specific enolase (NSE), microtubule-associated protein-2 (MAP-2) and glial fibrillary acidic protein (GFAP) were detected using immunocytochemical staining in each group. NSE, MAP-2 and GFAP expression was determined using Western-blot assay. RESULTS AND CONCLUSION: 12 and 24 hours following induction, BMSCs were positive for MAP-2, GFAP and NSE in the conditioned medium, bFGF and serum-free medium groups, but negative in the negative control group. Compared with the negative control group, MAP-2 expression was significantly enhanced in the conditioned medium, bFGF and serum-free medium groups 24 hours following induction (P < 0.05), and the increased range was significantly greater in the conditioned medium group compared with other two groups (P < 0.05). No significant difference in NSE and GFAP expression was detected in the conditioned medium, bFGF and serum-free medium groups. Results suggested that hippocampal neuron conditioned medium can in vitro induce the differentiation of rat BMSCs into neuron-like cells and glial cell-like cells. Compared with the bFGF medium and serum-free medium, positive rate was greatest in the hippocampal neuron conditioned medium-induced neurons and glial cells.