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[PDX-1 gene transfection and expression in bone marrow stromal cells-derived nestin-positive cells]
Hai-lan Wang1, Ming-xin Pan, Hui-ying Zhang
1Department of General Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
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.
Conclusion:
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.
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