绿色荧光蛋白转基因小鼠来源肌卫星细胞的体外培养及体内示踪

In vitro culture and in vivo tracking of muscle satellite cells derived from green fluorescent protein transgenic mouse

目的：从绿色荧光蛋白转基因小鼠中分离培养肌卫星细胞（MSCs）并进行体内示踪。方法：利用差速贴壁结合克隆分选方法，分离了MSCs，并于体外进行培养传代、鉴定及分化。检测所获MSCs的生长曲线及细胞周期并与来源于野生型鼠的同代MSCs进行比较。将绿色荧光蛋白标记的MSCs注射到裸鼠胫前肌，于注射后当时、注射后1周、2周、3周和4周利用二维荧光成像平台进行体内示踪。结果：MSCs被成功分离、传代及鉴定。来源于绿色荧光蛋白（GFP）标记或未标记小鼠MSCs的生长曲线、细胞周期及肌原性分化等无差别。MSCs注射后4周内可以动态观察到注射部位的绿色荧光信号并获得组织学证实。结论：来源于GFP转基因小鼠的MSCs在生长和增殖特性上与未转基因来源的MSCs相似，在体内可以通过二维荧光成像平台进行可靠的、无创性的示踪。

Objective： To isolate and culture muscle satellite cells （MSCs） from the green fluorescent protein （GFP） transgenic mouse （C57BL/Ka-13aetin-EGFP）, and then perform in vivo tracking. Methods： Through differential adhesion method combined with clones isolating, MSCs were obtained, then cultured, passaged, and identified. Growth curve, cell cycle of MSCs was tested and compared to counterparts of nontransgenic MSCs. Planar fluorescence imaging station （PFIS） was applieated to track GFP labeled MSCs 1, 2, 3 and 4 weeks after being injected into the tibialis anterior muscle of nude mouse immediately, and histological observation was performed. Results： MSCs were isolated, passaged and identified successfully. No significant difference was discovered in growth curve or cell cycle between GFP lahled MSCs and non-labled counterparts. Local green fluorescence was dynamically observed and recorded through PFIS throughout the whole 4 weeks after MSCs injections and was confirmed histologically. Conclusion： MSCs isolated from GFP transgenic mice were proved to be similar to their counterparts from non-transgenic ones in characters of growth and proliferation, and can be tracked in vivo through PFIS in a reliable and non-invasive way.