Effect of EGFP gene transfection on the cell cycle distribution of primary cultured human chondrocytes
- VernacularTitle:增强型绿色荧光蛋白基因转染对原代培养的人软骨细胞细胞周期的影响
- Author:
Xun JIANG
;
Yaoying ZENG
;
Xianhui HE
;
Lihui XU
;
Jingfang DI
;
Zheng FENG
;
Jingxian ZHAO
;
Qing WANG
;
Tong WANG
;
Jianbo SHI
- Publication Type:Journal Article
- Keywords:
Engineering, biomedical;
Chondrocytes;
Green fluorescent protein;
Gene transfer techniques;
Cell cycle
- From:
Chinese Journal of Pathophysiology
1989;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To investigate the effect of enhanced green fluorescence protein (EGFP) gene transfection on the cell cycle distribution of primary cultured human chondrocytes in order to establish a tracking method of cultured human nasoseptal chondrocytes. METHODS: pEGFP-N1 plasmid was amplified in E.coli, and purified by high purity kit. Primary cultured human chondrocytes,which were initially obtained from the nasoseptal cartilage, were cultured in vitro and transferred with pEGFP-N1 by means of electroporation with Amaxa nucleofector device. Transfering process and transient expression were evaluated by laser scanning confocal microscope (LSCM), the transfer efficiency and the cell cycle distribution were evaluated by flow cytometry. RESULTS: There was significant expression of EGFP at 24 h after transferring. The transfection efficiency of pEGFP-N1 into primary cultured human chondrocytes reached 35 37% at 48 h. It didn't affect the process of cell adherance and had no effect on the cell cycle distribution. CONCLUSION: Primary cultured human chondrocytes, which were transfected with pEGFP, are alive in vitro, and the transferring process doesn't affect the cell cycle distribution. These results suggest that pEGFP-N1 is an ideal transient expression vector for primary cultured human chondrocytes and it might be a well tracer in construction tissue engineered cartilage.
