The labeling of 3T3-L1 preadipocyte cells with enhanced green fluorescent protein.
A cell model is desired for adipocyte differentiation investigation and for high-throughput screening of anti-obesity and anti-diabetes molecules from chemical resources due to the world wide epidemic of obesity and diabetes. In order to establish such a cell model, a plasmid of pPPARgamma2-promoter-EGFP was constructed by inserting a 660bp sequence of mouse PPARgamma2 promoter into the Ase I and Kpn I sites of pEGFP-N3 and transferred into 3T3-L1 preadipocyte cells. The cells were induced to differentiate and the expression of PPARgamma2 was detected by the microscopic observation of EGFP and by RT-PCR assays. The results showed that the EGFP gene expression patterns were similar to that of pPPARgamma2's, which indicated that the EGFP gene was transferred into the mouse 3T3-L1 preadipocyte cells, and its expression was under the control of pPPARgamma2 promoter. RT-PCR assays showed that the EGFP expression authentically represented the stable expression of PPARgamma2. In conclusion, a preadipocyte cell line expressing EGFP under the control of the promoter of adipocyte-specific expression gene PPARgamma2 was generated. The cell line provides a powerful approach for the research of adipocyte differentiation and for the high-throughput screening of anti-obesity and anti-diabetes chemicals.
Li
Cheng-Jian
CJ
College of life Sciences, Peking University, Beijing 100871, China.
Cheng
Jun-Ying
JY
Zhang
Xiao-Lan
XL
Zhang
Chong-Ben
CB
D041721:3T3-L1 Cells; D017667:Adipocytes / Q000166:cytology*; D000818:Animals; D002454:Cell Differentiation; D049452:Green Fluorescent Proteins / Q000235:genetics*; D051379:Mice; D047495:PPAR gamma / Q000235:genetics*; D011401:Promoter Regions, Genetic; D013234:Stem Cells / Q000166:cytology*
D047495:PPAR gamma; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins