A study on transfecting green fluorescent protein gene to rat bone marrow mesenchymal stem cells.
- Author:
Zhi-yong LI
1
;
Lei LIU
;
Wei-dong TIAN
;
Xi-zhe CHEN
;
Zheng-bin YAN
;
Yun-feng LIN
Author Information
- Publication Type:Journal Article
- MeSH: Adenoviridae; genetics; Animals; Bone Marrow Cells; cytology; metabolism; Cell Differentiation; Cell Proliferation; Cells, Cultured; Genetic Vectors; Green Fluorescent Proteins; genetics; metabolism; Mesenchymal Stromal Cells; cytology; metabolism; Rats; Rats, Sprague-Dawley; Transfection
- From: Chinese Journal of Stomatology 2005;40(2):150-153
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSSD 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.
RESULTSAd-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.
CONCLUSIONSThe infected MSCs with Ad-GFP expressed GFP with much higher efficiency than liposome transfection, and maintained the same ability of proliferation and differentiation as non-infected MSCs. Transfection with Ad-GFP is a highly effective method for labeling MSCs.
