[PDX-1 gene transfection and expression in bone marrow stromal cells-derived nestin-positive cells].
OBJECTIVE
To 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.


METHODS
The 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.


RESULTS
Satisfactory 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.


CONCLUSIONS
The 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.
Wang
Hai-lan
HL
Department of General Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
Pan
Ming-xin
MX
Zhang
Hui-ying
HY
An
Jing
J
Gao
Yi
Y
D001854:Bone Marrow Cells / Q000378:metabolism*; D005822:Genetic Vectors; D018398:Homeodomain Proteins / Q000235:genetics*; D006801:Humans; D007381:Intermediate Filament Proteins / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D064231:Nestin; D010957:Plasmids; D017154:Stromal Cells / Q000378:metabolism*; D015534:Trans-Activators / Q000235:genetics*; D014162:Transfection / Q000379:methods*
D018398:Homeodomain Proteins; D007381:Intermediate Filament Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; D064231:Nestin; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein