A study on transfecting green fluorescent protein gene to rat bone marrow mesenchymal stem cells

• Published in: Zhonghua kouqiang yixue zazhi Vol. 40; no. 2; p. 150
• Main Authors: Li, Zhi-yong, Liu, Lei, Tian, Wei-dong, Chen, Xi-zhe, Yan, Zheng-bin, Lin, Yun-feng
• Format: Journal Article
• Language: Chinese
• Published: China 01.03.2005
• Subjects: Adenoviridae - genetics; Animals; Bone Marrow Cells - cytology; Bone Marrow Cells - metabolism; Cell Differentiation; Cell Proliferation; Cells, Cultured; Genetic Vectors; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - metabolism; Rats; Rats, Sprague-Dawley; Transfection
• ISSN: 1002-0098

To study an efficient method to transfect green fluorescent protein gene (GFP) to rat bone marrow mesenchymal stem cells (MSCs) and to determine the biological properties and differentiation potency of transfected MSCs. SD rats' bone marrow MSCs were separated and purified in vitro. After subculture and expansion, MSCs infected with Adenoviral vector (Ad-GFP) or transfected with liposome were observed, and their transfection efficiency was assessed with flow cytometry. The MSCs expressing GFP gene were induced to differentiate to osteoblast, and non-transfected MSCs were set as control. Ad-GFP delivered GFP gene with high efficiency to rat MSCs. (41.3 +/- 1.4)% of MSCs infected with Ad-GFP expressed GFP gene, which was much higher than the control (12.5%). Expression of GFP gene of infected MSCs maintained stable from 1 to 6 weeks after infection. Infected MSCs possessed the same alkaline phosphatase activation as non-infected MSCs, and formed mineralized mouldes. The infected MSCs with Ad-GFP expressed GFP w