Effects of hypoxia on the differentiation of human bone marrow mesenchymal stem cells into neural cells

LIU Li-jin; SHI Jian-ming; YIN Chang-chang

doi:10.3969/j.issn.1001-9448.2017.20.001

Abstract：Objective To investigate the effect of hypoxia on the differentiation of human bone marrow mesenchymal stem cells (hBMSCs) into neural cells.Methods The hMSCs (Passage 2) were purchased from Cyagen Biosciences Inc and identified by flow cytometry at Passage 4.The α-MEM medium containing 1％ FBS was used as the basal inducing medium.According to culture conditioning,they were divided into 4 groups,normoxia group,hypoxia group (3％ O2),normoxia-induced group (adding 20 ng/mL EGF and 20 ng/mL bFGF),and hypoxia-induced group (adding 20 ng/mL EGF and 20 ng/mL bFGF under the condition of 3％ O2).RT-PCR and Western blot were used to assess the mRNA and protein levels of the neuron markers,including NSE and MAP-2,and GFAP for glial marker after continuous inducing of 3 days.Results The identification results of flow cytometry showed that the expression of bonemarrow stroma marker CD105,CD90 and CD29 were 95.8％,98.2％ and 95.1％,respectively;while the expression of CD19,as the marker of hematopoietic cell,was 0.2％.Compared to normoxia group,the mRNA and protein levels of NSE,MAP-2 and GFAP were similar to hypoxic-group,while those were significantly increased both in normoxia-induced group and hypoxia-induced group (P ＜ 0.05);between which,they were significantly higher in hypoxia-induced group (P ＜ 0.05).Conclusion Hypoxia promotes the induction of EGF combined with bFGF on differentiation of hBM-SCs into neural cells and neurogliocytes.