小鼠3T3-L1前脂肪细胞系的特异性标记(英文)

用增强绿色荧光蛋白特异性标记小鼠 3T3 L1前脂肪细胞系 .构建paP2 promoter EGFP载体 ,电穿孔转染小鼠 3T3 L1前脂肪细胞 ,显微荧光观察和RT PCR确认aP2基因的内源表达 .EGFP基因转入 3T3 L1前脂肪细胞 ,观察到细胞分化过程中EGFP表达和脂肪积累 .RT PCR分析表明 ,EGFP代表了稳定而真实的aP2基因的内源性表达 .建立了由脂肪组织特异表达基因aP2的表达控制的EGFP标记的小鼠 3T3 L1前脂肪细胞系 ,目前尚未见用同样方法对前脂肪细胞进行特异性标记 .该细胞系将为脂肪细胞分化机理研究以及为抗肥胖症和抗糖尿病药物筛选提供有力工具 .

Specific Labeling of Mouse 3T3-L1 Preadipocyte Cell Line with Green Fluorescent Protein

A vector of paP2-promoter-EGFP was constructed and introduced into mouse 3T3-L1 preadipocyte cells, a cell line derived from mouse Swiss3T3 cells that were isolated from mouse embryo, to make the cells labelled with enhanced green fluorescent protein (EGFP) whose expression was controlled by the promoter of adipose-specific gene aP2. The cells were then induced to differentiate and the expression of aP2 was detected by EGFP-microscopy and RT-PCR assays. The EGFP gene was transferred into the mouse 3T3-L1 preadipocyte cells, and EGFP expression and lipid accumulation were observed during differentiation. The expression of aP2 was stable and similar to the expression of EGFP. A preadipocyte cell line expressing EGFP was obtained under the control of the promoter of adipocyte-specific expression gene aP2, and the preadipocyte cell line was specifically labelled. The cell line provides a powerful approach for the research of adipocyte differentiation and for the screening of anti-obesity and anti-diabetes drugs.