Construction Of Lentiviral Vector LIF And Stable Expression Of The Vector In The Bone Marrow Derived Mesenchymal Stem Cells

Purpose:The LIF gene in vitro transfection transfection on bone marrow mesenchymalstem cells transfected successfully tested to determine LIF can be stably expressed in bonemarrow after transfection of mesenchymal stem cells to promote peripheral nerveregeneration, reducing scar.Method:(1) of rat bone marrow mesenchymal stem cells separation and extraction,primary culture and phenotypic identification, by density gradient centrifugation combinedadherent vitro cultured rat bone marrow mesenchymal stem cells were observed underinverted phase contrast microscope morphological and draw its growth curve, measuring thesurface markers of mesenchymal stem cells were identified by flow cytometry.(2) LIF slowbuild viral vector packaging.(3) LIF lentiviral vector in bone marrow mesenchymal stemcells transfected with the ELISA test LIF gene and protein expression levels.Result:(1) bone marrow mesenchymal stem cells isolated from the success of theweek-old rat bone marrow blood, the microscope showed fibroblast-like growth can be madeto adipogenic, osteogenic cells transformed by flow cytometry, which surface markers CD44,CD90expression was positive, while CD34, CD45expression were negative;(3) proteinswere identified by ELISA, the purpose of LIF gene was stably expressed in rat bone marrowmesenchymal stem cells.Conclusion:(1) bone marrow mesenchymal stem cells isolated from the success of theweek-old rat bone marrow blood, the microscope showed fibroblast-like growth can be madeto adipogenic, osteogenic cells transformed by flow cytometry, which surface markers CD44,CD90expression was positive, while CD34, CD45expression were negative;(3) proteinswere identified by ELISA, the purpose of LIF gene was stably expressed in rat bone marrowmesenchymal stem cells.