Study Of Basic Fibroblast Growth Factor And Epidermal Growth Factor And Complex Factors Inducing Differentiation Of Rat Bone Mesenchymal Stem Cells Into Fibroblast

Objective:Comparing the role of the differents concentration of EGF and bFGF and complex factors inducing differentiation of Rat bone mesenchymal stem cells into Fibroblast. Research into growth factor-induced proliferation and differentiation of BMSCs best type and concentration.as to provides an ideal seed cells for the ligament tissue engineering.Methods:1 Whole bone marrow culture adherent, cell growth was observed under inverted microscope proliferation, determination of BMSCs during 1-8 days in primary with MTT, immunohistochemical detection BMSCs.2 Different concentrations on 2ng/ml,20ng/ml of bFGF, EGF and the combination of the concentration of the compound factor inducing medium instead of normal culture medium on the third generation of bone marrow mesenchymal stem cells induced differentiation, MTT test different concentrations after the induction of cell proliferation, immunohistochemistry for Laminin antigen detection.Results:1 Observed under an inverted microscope, bone marrow mesenchymal stem cells adherent after 24h, the shape of the cells changed from circular to spindle ,nuclei obviously, relatively large cytoplasm.2 After passage 2 cells were spindle, a small number of the polygon, cell fusion, or after the cells were vortex radial arrangement.3 MTT results showed that: Bone marrow mesenchymal stem cells were "S" shaped curve of growth. Primary cells into the logarithmic phase the next day, entered the fourth day of passage cells in logarithmic growth phase.4 Immunohistochemistry showed that bone marrow mesenchymal cell surface antigen CD34, Laminin expression was detected, Vimatin and expression ofα-SMA positive. After induction, the cells became positive for Laminin expression.5 MTT (7,14 days) and immunohistochemistry results:①All different concentrations of single and composite factor bFGF or EGF can induce bone marrow mesenchymal stem cells successfully differentiate into fiber cells;②The same concentration of EGF induced group and the group of bFGF group Comparison between the induced no significant difference;③Low concentration induced group 2ng/ml bFGF group, 2ng/ml EGF group, 2ng/ml bFGF + 2ng/ml EGF compared with the control group no significant difference;④High concentration induced group 20ng/ml bFGF group, 20ng/ml EGF group, 20ng/ml bFGF + 2ng/ml EGF group, 2ng/ml bFGF + 20ng/ml EGF and 20ng/ml bFGF + 20ng/ml EGF proliferation than the control group significantly accelerated, induced to differentiate into fibroblasts strongest, but high concentration no significant differences between groups.Conclutions:1 Whole bone marrow adherent culture method successfully cultured in vitro bone marrow-derived mesenchymal stem cells and obtain a higher purity.2 Two different concentrations of single and composite factor bFGF or EGF can induce bone marrow mesenchymal stem cells differentiate into fiber cells successfully, 20ng/ml is the ideal concentration inducing differentiation of rat bone mesenchymal stem cells into fibroblast.3 The same concentration bFGF and EGF to promote proliferation of BMSCs differentiation is basically the same.4 The induced effect of combined bFGF and EGFor used alone had no significant enhancement to BMSCs.