[Application of tracing technique of labeling bone marrow mesenchymal stem cells with green fluorescent protein in
Caixia Xu1, Peng Xiang, Rui Chen
1Biomaterials Research Insititute, College of Materials Science and Engineering, South China University of Technology, Ministry of Education, Guangzhou 510640, China. xucx@scut.edu.cn
Abstract:
This study was designed to label rat bone marrow mesenchymal stem cells (rBMSCs) with humanized renillar green fluorescent protein (hrGFP) for exploring the detectable effects of hrGFP on the construction of tissue-engineered bone in vitro. The hrGFP expression plasmid was packed into lentivirus by 293FT cells and transduced into rat BMSCs. After transduction, 81.3% rBMSCs successfully expressed green fluorescence. The hrGFP-rBMSCs were statically loaded on Bioglass-Collagen-Hyaluronic Acid-Phosphatidylserine (BG-COL-HYA-PS) scaffold in complete L-DMEM medium or osteogenic medium for 14 days. At 6h, a number of cells expressing green fluorescence can be observed by fluorescent microscopy. At day 7 and 14 after co-culture, the number of cells on the scaffold gradually increased. After 14 days for osteogenic induction, the hrGFP-rBMSCs and the interior of scaffold can be detected the expression of type I collagen. The results demonstrate that hrGFP labelling technique can detect visualizedly and effectively cell adhesion, proliferation and osteogenic differentiation in the construction of tissue-engineered bone in vitro.
