新生小鼠肌源性干细胞的生物学特性及表型研究

Isolation and biological characteristics of newborn mice muscle-derived stem cells

目的通过体外新生小鼠肌源性干细胞(MDSCs)的培养,观察鼠肌源干细胞的生物学特性及表型。方法采用胶原酶Ⅺ和胰蛋白酶分步消化肌肉,后采用差速贴壁分离技术纯化获得MDSCs。用倒置显微镜观察细胞形态,测定生长曲线,分析MDSCs的增殖能力,采用Sca-1、CD34和Desmin免疫细胞化学染色对细胞进行鉴定,初步确定细胞表型。结果经过两步消化和差速贴壁后,成功培养出高纯度的肌源性干细胞,该细胞呈Sca-1、CD34和Desmin阳性。结论可通过体外原代培养获得高纯度的肌源性干细胞,细胞具有良好的增殖能力,是适用于组织工程和基因治疗研究的一种新型种子细胞。

Objective To explore the feasibility of isolation and characteristics of the muscle-derived stem cells（MDSCs） of newborn mice.Methods To get purified MDSCs,newborn mice muscle were digested to single cells by collagenase and trypsin digestion,then were separated through differential adhesion.MDSCs were observed morphology under inverted microscope,reproductive activity was analyzed by determining growth curves and cells were identified by immunostaining with anti-Sca-1,CD34 and Desmin antibodies.Results Highly purified MDSCs were harvested and showed Sca-1（＋）、CD34（＋） and Desmin（＋）.Conclusion MDSCs can be isolated and cultured in vitro.MDSCs possess good proliferative ability and is adaptable for tissue engineering and gene therapy.