Objective       To determine the effects of bone morphogenetic protein 2 (BMP2) and glial cell line-derived neurotrophic factor (GDNF) on inducing migration

proliferation and differentiation of enteric nervous crest stem cells (ENCSCs)  as well as the signal transduction pathways involved.        Methods       (1) ENCSCs were obtained from the fetal intestine of SD rats by mechanical separation and enzyme digestion. (2) Immunocytochemical staining of nestin

P75

TUJ1

GFAP and SMA were performed for identifying whether the cells isolated and cultured were ENCSCs. (3) ENCSCs were cultured with or without GDNF and/or BMP2. Cell proliferation and migration was assessed by immunobiochemical staining based on 3-D culture model.        Results       (1) ENCSCs were successfully obtained from fetal intestine of rats. (2) ENCSCs could be passaged repeatedly

express nestin and P75

and differentiate to TUJ1-positive neurons

GFAP-positive glial cells

and SMA-positive smooth muscle cells. (3) Compared with control group

BMP2 could promote ENCSCs proliferation and migration (P0.05) and GDNF could enhance the biological effects (P0.05).        Conclusion       ENCSCs have the ability of proliferation and multi-potential differentiation. Proliferation and migration of ENCSCs are promoted by BMP2

while GDNF can enhance the biological effects.