Transfection and expression of leptin gene in human placental mesenchymal stem cells

VernacularTitle:瘦素基因在人胎盘间充质干细胞中转染与表达
Author: Zhenwei ZHU ; Chunli LIU ; Bowei WANG ; Yifu CHEN ; Zhanyi WANG ; Bei GAO ; Zhihui LIU
Publication Type:Journal Article
Keywords: Eukaryotic expression vector; Human placental mesenchymal stem cells; Leptin
From: Journal of Practical Stomatology 2014;(4):487-491
CountryChina
Language:Chinese
Abstract: Objective:To construct a eukaryotic expression vector of leptin gene,and to transfect it into human placental mesenchy-mal stem cells(HPMSCs)and appraise its expression.Methods:Primers were designed and the leptin gene was obtained by RT-PCR from human adipose tissue.The aimed segments were inserted into the eukaryotic expression vector pIRES2-EGFP,plasmid pIRES2-EGFP-LEP was constructed and identified by restricted enzymatic resection,and then transfected it into HPMSCs by liposome.The ex-pression of leptin in the transfected cells was detected by RT-PCR and Western blotting,the multi-differentiation potential of the cells was indentified.Results:The length of specific fragment was 500 bp,the recombinant plasmid pIRES2-EGFP-LEP presented 5.3 kb and 500 bp bands by restriction enzyme digestion.Leptin gene was expressed in transfected HPMSCs and the transfected HPMSCs maintained multi-directional differentiation potential.Conclusion:The eukaryotic expression vector of leptin can be transfected and ex-pressed in HPMSCs.