High expression of PDX-1 in bone marrow mesenchymal stem cells mediated by superfect / 中国病理生理杂志
Chinese Journal of Pathophysiology
; (12)1986.
Article
in Zh
| WPRIM
| ID: wpr-528865
Responsible library:
WPRO
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.
Full text:
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Index:
WPRIM
Language:
Zh
Journal:
Chinese Journal of Pathophysiology
Year:
1986
Type:
Article