A stem cell-based tool for small molecule screening in adipogenesis.
Techniques for small molecule screening are widely used in biological mechanism study and drug discovery. Here, we reported a novel adipocyte differentiation assay for small molecule selection, based on human mesenchymal stem cells (hMSCs) transduced with fluorescence reporter gene driven by adipogenic specific promoter--adipocyte Protein 2 (aP2; also namely Fatty Acid Binding Protein 4, FABP4). During normal adipogenic induction as well as adipogenic inhibition by Ly294002, we confirmed that the intensity of green fluorescence protein corresponded well to the expression level of aP2 gene. Furthermore, this variation of green fluorescence protein intensity can be read simply through fluorescence spectrophotometer. By testing another two small molecules in adipogenesis--Troglitazone and CHIR99021, we proved that this is a simple and sensitive method, which could be applied in adipocyte biology, drug discovery and toxicological study in the future.
Qin
Jie
J
Center for Stem Cell Biology and Tissue Engineering, Sun Yat-Sen University, Guangzhou, Guangdong, People's Republic of China.
Li
Wei-Qiang
WQ
Zhang
Li
L
Chen
Fei
F
Liang
Wen-Hua
WH
Mao
Frank Fuxiang
FF
Zhang
Xiu-Ming
XM
Lahn
Bruce T
BT
Yu
Wei-Hua
WH
Xiang
Andy Peng
AP
D017667:Adipocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D050156:Adipogenesis / Q000187:drug effects*; D002454:Cell Differentiation / Q000187:drug effects; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism
D049452:Green Fluorescent Proteins