小鼠成体心脏干细胞的分离和鉴定

The isolation and identification of adult cardiac stem cells in mice

目的建立稳定的心脏干细胞提取方法,为进一步研究心脏干细胞生物学特性及临床应用奠定基础。方法通过机械法剪碎组织,利用胶原酶Ⅱ消化后过细胞筛,再经流式细胞术根据细胞表面标志以及免疫荧光染色法进行鉴定,无菌分选得到细胞,体外培养。结果通过酶消化过筛法获得单细胞悬液,经流式细胞术细胞表面标志鉴定出Lin-CD45-Sca-1+细胞,无菌分选获得高纯度细胞可进行体外培养。结论本实验建立的小鼠成体心脏干细胞提取方法,可获得足够量的Lin-CD45-Sca-1+细胞用于实验研究。

Objective To establish a stable method of isolating cardiac stem cells for further study of cardiac stem cells biological characteristics and clinical application. Methods The stem cells were isolated by the following methods： mechanical shear cardiac tissue and collagenaseⅡ digestion. The marker on the surface of cardiac stem cells were recognized by the flow cytometry and immunofluorescence assays. And germ-free sorting cells were cultured in vitro. Results The single cells were obtained by enzyme digestion and the Lin^-CD45^-Sca-1^＋ cells were identified by the flow cytometry. Conclusion The cardiac stem cellsisolstion method were established,which could provide enough Lin^-CD45^-Sca-1^＋ cells used for scientific research. This study provided a technical support for treating heart disease with stem cells.