Differentiation Of Mesenchymal Stem Cells Derived From Human Umbilical Cord Into Neuro-like Cells

Objective:To isolate and culture human umbilical cord mesenchymal stem cells (hUCMSCs) in vitro. To explore an ideal method to induce the differentiation from hUCMSCs into neuron-like cells and provide some evidence for the transplantation of hUCMSCs for spinal cord injure, different cytokines were used to induce the differentiation.Methods:The hUCMSCs were isolated from the human umbilical cord digested with collagenaseâ…¡. Morphologic appearance of hUCMSCs was observed by an atomic force microscopy, and the hUCMSCs was verified with flow cytometry. All of Passage 5 cells were divided into four groups at random? marked with A,B,C and D respectively. The differentiation of hUCMSCs were induced by using bFGF in group A, bFGF+BDNF in group B, and BHA+bFGF+BDNF in group C respectively while group D as a control group was cultured by DMEM-F12 with10%FBS. Two weeks later the expression of Nestin, neurofilament heave (NEFH) and glial fibrillary acidic protein (GFAP) were detected using real-time PCR kit and immunocytochemistry, and the morphologic changes of cells were observed by an atomic force microscopy.Results:Mesenchymal stem cells can be isolated and cultured from human umbilical cord by enzyme digestion. The passage 3 and 7 cells present a large expansive potentiality detected by MTT. HUCMSCs express CD29,CD44,CD105 but no CD34,CD45 and HLA-DR. After cultured with inducing medium for 2 weeks, the cells were successfully induced into neuro-like cells. The appearance of the cells have great change.The induced hUCMSCs developed rounded cell bodies with multiple neurite-like extensions observed by an atomic force microscopy. The result of immunocytochemistry and RT-PCR showed that Nestin was positive in A,B and C groups, NEFH was positive in A and B groups, but in the four groups the GFAP was negative. The difference of the expression of Nestin and NEFH among the induced groups was significantly (P<0.05), analysed by SPSS 13.0.Conclusions:Mesenchymal stem cells can be isolated and cultured from human umbilical cord by enzyme digestion in vitro, and all the hUCMACs present stable biological properties. Moreover, hUCMSCs can be induced and differentiated into neuro-like cells in vitro via bFGF combined with BDNF.