Construction of tissue engineering adipose with human umbilical cord mesenchymal stem cells transfected by insulin gene and silk fibroin scaffold in vitro
10.3760/cma.j.issn.1671-0290.2012.04.012
- VernacularTitle:胰岛素基因转染的人脐带间充质干细胞与丝素蛋白支架构建组织工程脂肪的研究
- Author:
Jun TANG
;
Yi LIU
;
Bin XU
- Publication Type:Journal Article
- Keywords:
Tissue engineering adipose;
Human umbilical cord mesenchymal stem cells;
Silk fibroin;
Insulin;
Gene transfection
- From:
Chinese Journal of Medical Aesthetics and Cosmetology
2012;18(4):277-281
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the construction of tissue engineering adipose in vitro with silk fibroin scaffold and human umbilical cord mesenchymal stem cells (hUCMSCs) transfected by recombinant human insulin gene lentivirus.Methods hUCMSCs infected with recombinant lentiviral pLenti6.3-insulin-IRES-EGFP (Group A) by the best MOI=10 were seeded in silk fibroin scaffold; hUCMSCs transfected by EGFP gene (Group B)was regarded as negative control; and then the cell-scaf-fold complexes were cultured in adipogenic differentiation medium. MTT test was used to detect whether recombinant lentiviral affected the growth and proliferation of hUCMSCs and the growth activity of hUCMSCs seeded in silk fibroin.Results After 5-7 days for adipogenic culture,the numbers of fat-like cells in group A were significantly more than those in group B (P =0.007,P＜0.01).RTPCR results showed that the expression of PPARγ-2 in group A was much stronger than that in group B.MTT test showed that there was no significant difference in optical density (A)at each time between transfected group and nontransfected group (P=0.056,P＞0.05).And there was also no significant difference in optical density (A)between cell group and cell-scaffold group (P =0.066,P＞0.05).Conclusions Insulin gene could obviously promote hUCMSCs getting into adipose,and carrying recombinant human insulin gene lentiviral vector transfection of hUCMSCs and silk fibroin scaffolds could effectively construct tissue engineering adipose in vitro.
