Cationic liposome-mediated enhanced green fluorescent protein plasmid transferred into skeletal muscle satellite cells

doi:10.3969/j.issn.2095-4344.2013.46.013

Abstract

Abstract:

BACKGROUND: Skeletal muscle satellite 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 cell engineering.
OBJECTIVE: To establish a thrifty, convenient culture procedure and create a simple, efficient method to transfect skeletal muscle satellite cells, and investigate genetic expression after the transfection for cellular cardiomyoplasty.
METHODS: Skeletal muscle satellite 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 satellite 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: Satellite cells were isolated, cultured and transfected successfully. 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 gradually after one week. The expression still could be observed two weeks latter. The enhanced green fluorescent protein plasmid conducted by cationic liposome could be transfered into skeletal muscle satellite cells efficiently. The transfection efficiency was correlated closely to the ratio of plasmid and lipofectamine. The change of target gene expression depended on time.

Key words: satellite cells, skeletal muscle, plasmids, green fluorescent proteins, transfection, cations, liposomes

CLC Number:

R318

Xu Zhi-feng, Li Jing-lai, Han Zhen, Feng Gang, Ren Ming-ming. Cationic liposome-mediated enhanced green fluorescent protein plasmid transferred into skeletal muscle satellite cells[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(46): 8056-8061.

Figures/Tables 5

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