Ad5-enhanced green fluorescent protein versus rAAV2-enhanced green fluorescent protein transfecting adipose-derived mesenchymal stem cells
10.3969/j.issn.1673-8225.2010.14.024
- VernacularTitle:Ad5-增强型绿色荧光蛋白和rAAV2-增强型绿色荧光蛋白转染脂肪间充质干细胞的对比
- Author:
Xiaohong YUAN
;
Rongze AN
;
Zhaojie WANG
;
Enuo JIA
;
Xinwen QI
;
Jinping CHEN
;
Jin YANG
;
Fanfan LIU
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2010;14(14):2588-2590
- CountryChina
- Language:Chinese
-
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 stabry.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.
