Impact of activated schwann cells-derived neuronotrophins on differentiation of human umbilical cord blood mesenchymal stem cells into neurogenic cellsin vitro

Abstract

Objective To observe the capability of human umbilical cord blood mesenchymal stem cells (HUCBMSCs) to differentiate into neurogenic cells in vitro under the impact of activated Schwann cells (ASCs)-derived neuronotrophins.Methods ASCs were cultured by using the dual-enzyme digestion method combined with mechanical separation.HUCBMSCs were cultured through isolating HCMNCs by Ficoll' s way and then plastic-adherent culture.Two methods were used for HUCBMSCs culture:all-trans retinoic acid + bFGF + EGF ( control group),1/2 DMEM + ASCs culture ( medium after ASCs cultured for 2 days,experimental group).Immunohistochemistry,Western blotting,and semi-quantitative and realtime polymerase chain reaction (PCR,NF200,GFAP,MBP) quantitative analysis were used to evaluate neurogenic differentiation of HUCBMSCs.Results The results of immunohistochemistry staining,such as NF200,GFAP and MBP,confirmed that HUCBMSCs could differentiate into neurogenic cells under the effect of ASCs and RA + bFGF + EGF.The statistical analysis of Western blotting and RT-PCR ( NF200,GFAP,MBP ) indicated that there existed significant difference between two groups.Conclusion HUCBMSCs can proliferate and differentiate into neurogenic cells under the effect of numerous neurotrophic factors from ASCs in vitro. Key words: Umbilical cord blood mesenchymal stem cells; Schwann cells; Neural repair; Differentiation