Neuronal differentiation effects of vascular endothelial factor on bone marrow stromal cells

BACKGROUND: Studies have demonstrated that bone marrow stromal cells (BMSCs) undergo neuronal differentiation under certain in vitro conditions. However, very few inducers of BMSC differentiation have been used in clinical application The effects of vascular endothelial growth factor (VEGF) on in vitro neuronal differentiation of BMSCs remain poorly understood OBJECTIVE: To investigate the effect of VEGF on neuronal differentiation of BMSCs in vitro, and to determine the best VEGF concentration for experimental induction. DESIGN, TIME AND SETTING: In vitro comparative study was performed at the Central Laboratory and Laboratory of Male Reproductive Medicine, Shenzhen Hospital of Peking University from October 2008 to August 2009 MATERIALS: Recombinant human VEGF165 was purchased from Peprotech Asia, Rehovot, Israel Neuron-specific enolase (NSE) was purchased from Beijing Biosynthesis Biotechnology, China METHODS: BMSCs were harvested from adult Sprague Dawley rats The passaged cells were pre-induced with 10 ng/mL basic fibroblast growth factor for 24 hours, followed by differentiation induction with 0, 5, 10, and 20 ng/mL VEGF, respectively MAIN OUTCOME MEASURES: Morphological changes in BMSCs prior to and following VEGF induction Expression of NSE following induction was determined by immunocytochemistry RESULTS: Shrunken, round cells, with a strong refraction and thin bipolar or multipolar primary and secondary branches were observed 3 days after induction with 5, 10, and 20 ng/mL VEGF However, these changes were not observed in the control group. At 10 days after induction, the number of NSE-positive cells was greatest in the 10 ng/mL VEGF-treated group (P < 0 05) The number of NSE-positive cells was least in the control group at 3 and 10 days post-induction (P < 0 05) Moreover, the number of NSE-positive cells was greater at 10 days compared with at 3 days after induction (P < 0 05) CONCLUSION: Of the VEGF concentrations tested, 10 ng/mL induced the greatest number of neuronal-like cells in vitro from BMSCsCell BiologyNeurosciencesSCI(E)中国科学引文数据库(CSCD)1ARTICLE9657-661