PDX-1 gene transfection and expression in bone marrow stromal cells-derived nestin-positive cells.
- Author:
Hai-lan WANG
1
;
Ming-xin PAN
;
Hui-ying ZHANG
;
Jing AN
;
Yi GAO
Author Information
- Publication Type:Journal Article
- MeSH: Bone Marrow Cells; metabolism; Genetic Vectors; Homeodomain Proteins; genetics; Humans; Intermediate Filament Proteins; metabolism; Nerve Tissue Proteins; metabolism; Nestin; Plasmids; Stromal Cells; metabolism; Trans-Activators; genetics; Transfection; methods
- From: Journal of Southern Medical University 2007;27(4):528-531
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo introduce the eukaryotic expression vector pEGFP-C1-PDX-1 into nestin-positive cell derived from bone marrow stromal cells by nucleofection and optimize the conditions for transfection.
METHODSThe recombinant plasmid was transfected into bone marrow stromal cells-derived nestin-positive cells with varied DNA quantities or the serum concentration in the medium. The expression of PDX-1 gene in the transfected cells was detected by RT-PCR.
RESULTSSatisfactory efficiency of transfection was achieved with the DNA quantity of 2-10 microg and medium serum concentration of 20%. PDX-1 expression was detected in the transfected cells by RT-PCR.
CONCLUSIONThe optimized transfection conditions result in enhanced efficiency of PDX-1 gene transfection into nestin-positive cells derived from bone marrow stromal cells, which may serve as the seed cells in tissue-engineering.
