Transfection of rat myoblasts with leuflvirus carrying autocrine motility factor gene

VernacularTitle:携带自分泌运动因子基因的慢病毒载体转染大鼠成肌细胞
Author: Pen LI ; Lan JIN ; Yi TIAN ; Zumeng YA
Publication Type:Journal Article
Keywords: Myoblast; Autocrine motility factor; Transfection; Feline immunodeficiencyvirus,feline
From: Chinese Journal of Trauma 2009;25(7):653-657
CountryChina
Language:Chinese
Abstract: Objective To explore a safe and high efficiency way of gene transfection of autocrine motility factor(AMF) in order to provide experimental basis for transplantation of myoblasts carrying AMF gone. Methods Sprague Dawley rat myoblasts were cultured, purified, proliferated and immunohisto-chemically verified. Then, the myoblasts were transfected with AMF and eGFP (enhanced green fluores-cent protein) gene by FIV (feline immunodeficiency virus). Fluorescence microscope and laser scanning confocal microscope were employed to detect eGFP so as to verify positive transfection rate. Expression of AMF was detected by immunohistochemical method. Results Myoblasts with 98% purity could he ob-tained after two weeks of primary culture and purification. Positive transfection rate reached 90.4% when MOI (multiplicity of infection) was 100 (P <0.01). The transfected AMF gene could express normally. Conclusions Explant culture is a proper way in rat myoblast culture. Meanwhile, AMF gene can he effectively transfected into rat myoblast and well expressed via FIV.