牛骨骼肌卫星细胞的分离鉴定和诱导分化

Isolation,Identification and Induced Differentiation of Bovine Skeletal Muscle Satellite Cells

本研究对牛骨骼肌卫星细胞进行体外分离培养、诱导分化和鉴定,采用胶原酶和胰酶联用的酶消化法分离肌卫星细胞,应用差速贴壁法进行纯化,观察卫星细胞及诱导分化后肌管的形态结构,并利用标志基因的反转录PCR(RT-PCR)和免疫荧光染色方法对分化前后细胞进行鉴定。结果显示,分离出的肌卫星细胞呈梭形生长,生长状态良好,RT-PCR和免疫荧光染色显示肌卫星细胞Pax7和MyoD呈阳性表达,纯化后的肌卫星细胞纯度大于93%;诱导分化后,卫星细胞融合生长,形成的肌管状态良好,分化标志基因MyoG和MHC呈阳性表达。本研究建立了一套从牛肌肉组织中分离和鉴定肌卫星细胞的方法,可以为肌肉的发育分化和肉牛肉质改良研究提供良好的细胞模型。

In this study, bovine skeletal muscle satellite cells were isolated and identified, muscle tissues were digested bycollagenase and trypsinase, then the satellite cells were purified and induced differentiate into myotubes. The muscle satellitecells and myotubes were observed, and detected by RT-PCR and immunofluorescent staining technology. The results showedthat the muscle satellite cells were in healthy morphology, and more than 93 percent of the cells expressed the marker genespaired box protein 7 （Pax7） and myogenic differentiation antigen （MyoD）. After myogenesis induction, satellite cells began toconvergence, and finally formed multinucleated myotubes expression of myogenin （MyoG） and myosin heavy chain （MHC）.Our results indicated that high purity skeletal muscle satellite cells were successfully isolated and identification, which couldprovide cell source for muscle development and bovine muscle quality improvement study.