High expression of PDX-1 in bone marrow mesenchymal stem cells mediated by superfect
- VernacularTitle:Superfect高效介导PDX-1基因在骨髓间质干细胞表达
- Author:
Jiping SUN
;
Yanjie JIA
;
Xiaoli WANG
;
Yujia YANG
- Publication Type:Journal Article
- Keywords:
Bone marrow mesenchymal stem cells;
Gene,pancreatic duodenal homeobox-1;
Diabetes mellitus
- From:
Chinese Journal of Pathophysiology
1986;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To construct a eukaryotic expression vector containing pancreatic duodenal homebox-1(PDX-1)and to elevate the expression efficiency of exogenous gene in rat bone marrow mesenchymal stem cells(MSCs).METHODS:Recombinant vector containing PDX-1 was constructed.Flow cytometry was used to identify the cell cycle of bone marrow mesenchymal stem cells(MSCs)cultured in vitro.Recombinant vector containing PDX-1 was transfected into bone marrow MSCs using superfect in medium.After being selected by G418,RT-PCR and Western blotting were used to investigate the expression of PDX-1 in MSCs.RESULTS:Restricted enzyme analysis and sequencing showed that PDX-1 gene segment was consistent with that in GenBank.Flow cytometry showed that there were about 85.9% cells at the cell cycle of G_0/G_1.The whole cells transfected emitted green fluorescence under flow cytometry.The efficiency of transfection was above 40%.RT-PCR and Western blotting demonstrated that there was expression of PDX-1 in transfected bone marrow MSCs.CONCLUSION:Recombinant vector containing PDX-1 was constructed successfully.Superfect mediated expression of exogenous gene in bone marrow MSCs in a high efficiency,and bone marrow MSCs containing exogenous gene are an ideal cells for gene therapy.
