Differentiation of Human Umbilical Cord Blood Mesenchymal Stem Cells into Neuron Like Cells in Vitro Induced by GM
MA Yan;GUO Jin-yao;LIU Lei;FAN Chun;General Hospital of Huabei Petroleum Administration;
Objective To study the monosialo four hexose ganglioside( GM1) induced human umbilical cord mesenchymal stem cells into neuron like cells and to explore the optimal concentration in vitro. Methods The umbilical cord,umbilical artery and umbilical vein were collected from the healthy fetus in full term pregnancy. The MSCs was obtained by enzyme digestion method and the cell phenotype was determined by flowcytometry. After amplification,the fourth generation cells were divided into A,B,C,D,E five groups. The first 4 groups were the experimental groups,E as control group. The experimental group cells were treated with different concentrations of GM1,group A 50μg / m L,B 100μg / m L group and C group150μg / m L group D 200μg / m L. Inducing medium was diluted by L-DMEM,the control group was only treated with L-DMEMmedium. The morphology of 5 groups was observed and compared every 60 min. At360 min,the expression of glial fibrillary acidic protein( GFAP) and neurofilament protein( neurofilaments protein H,NF-H) and neuron specific markers of microtubule-associated protein 2( MAP-2) was examined by immunohistochemistry,and calculate the rate of positive cells. Results Most of the primary cells were adherent at 9h after inoculation. The cells evolved from polygonal and diamond shapes to spindle shape,and then into spiral and radial shape. Expression of CD105 and CD73,but not HLA-DR,CD90,CD19,CD11 b,CD45,CD34,and P3 were detected by flowcytometry. The molecular markers of P5 and P10 on cell surface were detected by flowcytometry. At 360 th minutes after induction,the cells of the experimental group were found to be neuron like cells,which showed that the cell bodies were contracted into oval shape,extending out longer processes,and the bipolar cells were the majority. The expression of NF-H and MAP-2were detected in the experimental group,whereas GFAP was negative. NF-H and MAP-2 in C group( 150μg /m L) was at highest rate. Conclusion monosialo four hexose ganglioside,GM1 can induce umbilical cord mesenchymal stem cells to differentiate into neuron like cells. The concentration of 150μg / m L was the most appropriate induction dose.
【CateGory Index】： R329.2