[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.

CONCLUSION

Cationic lipid is an effective carrier for gene-modified cell transplantation therapy.