Construction of rat bone marrow mesenchymal stem cells modified with human hepatocyte growth factor gene
10.3760/cma.j.issn.0254-1416.2012.09.027
- VernacularTitle:人肝细胞生长因子基因修饰大鼠骨髓间充质干细胞系的构建
- Author:
Qun LIN
;
Lihua LEI
;
Caizhu LIN
;
Xianzhong LIN
;
Jianqing LIN
;
Huizhe ZHENG
;
Hongda CAI
;
Qing YANG
;
Youguang GAO
- Publication Type:Journal Article
- Keywords:
Hepatocyte growth factor;
Organisms,genetically modified;
Stem cells;
Bone marrow
- From:
Chinese Journal of Anesthesiology
2012;(9):1126-1129
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct F344 rat bone marrow mesenchymal stem cell line (MSC) modified with human hepatocyte growth factor (hHGF) gene.Methods Recombinant virus containing hHGF was obtained by transfecting the packaging cell line 293 FT with lentiviral vector pLV/EF1α-hHGF-IRES-eGFP.MSCs derived from F344 rat bone marrow were then tranfected with packed lentiviral vector.Purified MSCs expressing hHGF was obtained by screening culture with G418.MSCs and MSCs transfected with empty vector were used as control.The expression of hHGF protein was detected by Western blot (eGFP-MSCs).The hHGF-transfected MSCs were cultured in osteoblast-inducing culture medium and osteoblast phenotype was assayed by alizarin Red staining.The cells were also cultured in adipogenesis medium and stained with Oil Red O for identification.Results The expression of hHGF protein was significantly up-regulated in the hHGF-MSCs as compared with MSCs and eGFP-MSCs.hHGF-MSCs readily differentiated into mineralizing cells or adipocytes when incubated in differentiation medium.Conclusion A F344 rat MSC line that stably expresses HGF is successfully established.
