Study Of TGF-β1 And EGFP Electropration On Rabbit Bone Marrow Derived Mesenchymal Stem Cells In Vitro

Objective:To observe expression of collagenⅡ by transfecting growth factor-β1(TGF-β1) and EGFP gene into rabbit bone mesenchymal stem cells(BMSCs) in electropration.Method : 3 New Zealand white rabbits(1 week old)were used,the Tibia and femur were separated immediately after euthanized rabbits under strictly aseptic conditions,exposed the canal and BMSCs were cultured after washed bone marrow and digestion in vitro;Flow cytometry was used to detect cell phenotype;BMSCs were divided into three groups: TGF-β1 and EGFP electropration group,EGFP electropration group,blank control group;TGF-β1 and EGFP plasmid were trasfected into BMSCs through electroporation,transfection efficiency was detected by fluorescence microscop; immunohistochemistry and western blot method type Ⅱ collagen were used to detect expression of collagen Ⅱafter 14 days.Result:BMSCs can be isolated and cultured by whole bone marrow cells adherent culture,CD90 and CD44 is positive,CD31 and CD45 is negative by flow cytometry; Successfully transfected TGF-β1 to the BMSCs,the strong expression of collagenⅡ in TGF-β1 group cells(0.4870±0.0181)is showed by Immunocytochemical stain,which was compared with empty plasmid group(0.0858±0.0097)and the control group(0.0805±0.0098);and the difference was statistically significant( P<0.05). the strong expression of collagenⅡin TGF-β1 group cells(0.320 0±0.006 7)is showed by western blot, which was compared with empty plasmid group(0.201 2±0.007 4)and the control group(0.196 4±0.004 1),and the difference was statistically significant(P<0.05).Conclusion:The expression of collagen Ⅱcan increase by TGF-β1 plasmid electroporation into rabbit BMSCs,it can establish foundation for next certain research.