Cationic liposome-mediated enhanced green fluorescent protein plasmid transferred into skeletal muscle satellite cells
10.3969/j.issn.2095-4344.2013.46.013
- VernacularTitle:阳离子脂质体介导增强型绿色荧光蛋白质粒转染骨骼肌卫星细胞
- Author:
Zhifeng XU
;
Jinglai LI
;
Zhen HAN
;
Gang FENG
;
Mingming REN
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2013;(46):8056-8061
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:Skeletal muscle satel ite cells are totipotential stem cells with multi-directional differentiation potential, locate in skeletal muscle interstitium, have a certain tolerance to ischemia and hypoxia, and are important cells in stem cellengineering.
OBJECTIVE:To establish a thrifty, convenient culture procedure and create a simple, efficient method to transfect skeletal muscle satel ite cells, and investigate genetic expression after the transfection for cellular cardiomyoplasty.
METHODS:Skeletal muscle satel ite cells were isolated from rabbit thigh and cultured. Their growth curves were determined by CKK-8 method. Grouped by different proportions of the plasmid and liposome, skeletal muscle satel ite cells were transfered by the enhanced green fluorescent protein plasmid based on liposome. After transfection, the efficiency and character of target genetic expression was determined.
RESULTS AND CONCLUSION:Satel ite cells were isolated, cultured and transfected successful y. In suitable ratio of plasmid and liposomes, the transfection efficiency reached up to above 35%. The target protein was expressed within 12 hours after transfection, reached maximum in 48-72 hours and decreased gradual y after one week. The expression stil could be observed two weeks latter. The enhanced green fluorescent protein plasmid conducted by cationic liposome could be transfered into skeletal muscle satel ite cells efficiently. The transfection efficiency was correlated closely to the ratio of plasmid and lipofectamine. The change of target gene expression depended on time.
