Abstract:

BACKGROUND: Liposome has toxic side effect on cells, but introduction of exogenous genes into cells affects cell metabolism. Therefore, it is worthy to explore effects of liposome-mediated human bone morphogenetic protein-2 (hBMP-2) gene-transfected adipose-derived stem cells (ADSCs) on cell growth and proliferation.
OBJECTIVE: To investigate the effects of liposome-mediated hBMP-2 gene transfection on growth and proliferation of rabbit ADSCs.
METHODS: The rabbit ADSCs were extracted from the adipose tissue and subcultured in vitro. The fourth passage of ADSCs were collected and transfected by hBMP-2 gene and enhanced green fluorescent protein (EGFP) gene respectively. The cell morphology and their related growth pattern were continuously observed under the inverted microscope after transfection. At 48 hours after transfection, the transient transfection efficiency was measured by fluorescence microscopic counting. MTT assay was used to drawn the cell growth curve and to calculate the colony doubling time.
RESULTS AND CONCLUSION: At 48 hours after transfection, cell volume became large, with abundant cytoplasm, and some were round or oval. The transient transfection rate was about (18.0±0.42)% under fluorescence microscope. According to the cell growth curves drawn by MTT assay, the transfected cells grow at a slower rate compared with the untransfected cells, but both of them roughly had the same growth curve, like the shape of “S”, and little change in their overall trends. After transfection, the population doubling time was 55.51 hours in the hBMP-2 group, 53.58 hours in the EGFP group and 46.10 hours in the non-transfected group. There was no significant statistical difference (P > 0.05). Results have indicated that liposomes can mediate the hBMP-2 gene and EGFP gene transfected rabbit ADSCs and it has no effect on their growth and proliferation.