Differentiation of rat bone marrow stromal cells into enteric neurons and expression change of glial cell line-derived neurotrophic factor

Abstract

Objective To investigate differentiation conditions of bone marrow stromal cells (BMSCs) to enteric neuron in vitro and expression of glial cell line-derived neurotrophic factor (GDNF) and its receptor RET. Methods BMSCs were harvested from rats and cultured in DMEM supplemented with 10% fetal bovine serum (FBS). After 4 passages, BMSCs were induced with 10ng/mlGDNF and fetal gut culture medium (FGCM) for 7 days. The expression of neural specific enolase (NSE). vasoactive intestinal peptide (VIP) and nitric oxide synthase (nNOS) was examined with immunohistochemistry. The change of GDNF and RET mRNA expressed was assessed with RT-PCR and protein expression was quantified with western blot. Results BMSCs were cultured and purified in vitro. After 7 days of induction, a certain number of cells showed expression of NSE, VIP and nNOS by immunohistochemistry method in experimental group, while the cells showed no expression of VIP and NSE in control group. BMSC expressed low GDNF and no RET mRNA, while after induction, the expression of GDNF mRNA was highly increased and RET mRNA expressed. GDNF protein was expressed. Conclusions GDNF and FGCM induce differentiation of BMSCs into enteric neurons,and GDNF-RET signaling pathway may play an important role in enteric neural differentiation. Key words: Mesenchymal stem cells; Glial cell line-derived neurotrophic factor (GDNF) ; Culture media