Abstract:

BACKGROUND: There are reports concerning differentiation of adipose-derived mesenchymal stem cells (ADSCs) into chondrocytes using gene transfection technique. However, the transfection of adenovirus and adeno-associated virus into ADSCs is various. It is controversial whether adeno-associated virus (AAV) can transfect ADSCs.
OBJECTIVE: To observe the enhanced green fluorescent protein (EGFP) expression following Ad5-EGFP and rAAV2-EGFP transfection into ADSCs, and investigate the cell proliferation ability following transfection.
METHODS: ADSCs were isolated from the adipose tissue, which was from 6-month-old New Zealand albino rabbit back and neck by mechanical digestion and enzyme digestion, then ADSCs were cultured and amplified in vitro. ADSCs were infected with Ad5-EGFP and rAAV2-EGFP, and the EGFP expression was observed. A total of 2 μL sodium butyrate (1 mol/L) was added into the medium after rAAV2-EGFP transfection. MTT assay was used to detect the gene transfection effects on reproductive activity of ADSCs.
RESULTS AND CONCLUSION: ADSCs isolated and cultured in vitro were flat, long-spindle and amplified stably. The cell morphology was uniform. Many green fluorescent cells were observed in Ad5-EGFP and rAAV2-EGFP groups. Transfection efficiencies were about 88% and 83%. Adenovirus and adeno-associated virus vector can be transfected with ADSCs, and transfection efficiency is high. Adeno-associated virus needs sodium butyrate to increase its level of gene expression.