摘要
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.
出处
《吉林医药学院学报》
2015年第3期167-170,共4页
Journal of Jilin Medical University
基金
国家自然科学基金资助项目(31260285)
关键词
心脏干细胞
酶消化
分离
鉴定
cardiac stem cells
enzyme digestion
isolation
identification