[Plasmid transfection of rat bone marrow mesenchymal stem cells by cationic lipid for gene-modified cell transplantation therapy].
OBJECTIVE
To investigate the feasibility of gene transfection of bone marrow mesenchymal stem cell (BMSC) by cationic lipid.


METHODS
The relative optimal transfection condition was determined by scale transfection experiment in vitro and the transient transfection efficiency of enhanced green fluorescence protein (EGFP) gene for rat BMSC was determined with Lipofectamine2000 (LP2000). The relationship between cell cycle status and the expression of the gene was analyzed. The intensity and the ratio of EGFP gene expression versus time was determined by flow cytometry. In the in vivo study, the transgenic rat BMSC was injected into the myocardium of inbred rats, and the in vivo transcription of EGFP gene and the EGFP-expressing BMSC were traced in the myocardium after transplantation using reverse transcription-polymerase chain reaction and fluorescent microscopy, respectively.


RESULTS
EGFP gene transfection efficiency in BMSC was different under different transfection condition (DNA concentration and DNA: LP2000). Cationic lipid-mediated transfection demonstrated marked toxicity to BMSC. Cell cycle status restricted the expression efficiency of the gene introduced by cationic lipid. The EGFP expressing-BMSC and in vivo transcription of the EGFP gene could be detected in rat myocardium post implantation.


CONCLUSIONS
Cationic lipid is an effective carrier for gene-modified cell transplantation therapy.
Zhang
Peng
P
Ministry of Health Key Laboratory of Cardiac Regeneration, Fuwai Hospital, CAMS and PUMC, Beijing 100037, China.
Zhang
Hao
H
Hu
Sheng-Shou
SS
Chen
Li-Guo
LG
Wei
Ying-Jie
YJ
D000818:Animals; D030801:Animals, Genetically Modified; D001854:Bone Marrow Cells / Q000166:cytology*; D002412:Cations; D002478:Cells, Cultured; D004337:Drug Carriers / Q000494:pharmacology; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics; D008055:Lipids; D008081:Liposomes / Q000494:pharmacology; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D010957:Plasmids / Q000235:genetics*; D051381:Rats; D011917:Rats, Inbred Lew; D017207:Rats, Sprague-Dawley; D014162:Transfection
D002412:Cations; D004337:Drug Carriers; D008055:Lipids; D008081:Liposomes; D049452:Green Fluorescent Proteins