摘要
造血干细胞极为稀少,约占骨髓中所有有核细胞的0.01%。几十年来,如何从骨髓中分离造血干细胞一直是研究的焦点问题。流式细胞术在识别和分离造血干细胞及祖细胞中起着关键性的作用。三十年间,有多种不同的细胞标志物和分离手段用于小鼠骨髓中造血干细胞的分析和分选。该文对所有分离策略作了分类总结,并详细介绍了现在认可度最高且应用最广泛的造血干细胞的分离方法:SLAM家族分离策略。然而,仅仅对造血干细胞进行体外的流式分析并不能完整描述造血干细胞在体内的行为特征。谱系示踪技术弥补了这一不足。该文使用α-catulin;和Fgd5-CreERT2;Rosa26-Tomato小鼠详细说明了如何使用流式细胞仪来分析和量化造血干细胞中的报告荧光蛋白信号。
Hematopoietic stem cells represent a rare population with an estimated frequency of 0.01% of total nucleated cells in bone marrow.Isolating and separating HSCs from bone marrow has been the focus of intense investigation for decades.Flow cytometry has been critical in establishing methods to isolate and identify hematopoietic stem cells and their progenitors.For more than 30 years,researchers have been uncovering and combining emerging markers used for improving the purification of hematopoietic stem cells from mouse bone marrow.This article summarizes all the phenotypic markers and strategies used to purify HSCs and details the most acknowledged and widely adopted SLAM family isolation protocol.However,in vitro flow cytometric analysis of hematopoietic stem cells does not provide a true description of the behavior of HSCs in their native physiological state.The application of genetic lineage tracing in HSCs has bridged the gap.This article uses α-catulin;and Fgd5-CreERT2;Rosa26-Tomato mice to detail instructions on how to use flow cytometry to analyze and quantify reporter fluorescent protein signals in HSCs.
作者
陈琳
李晶晶
周波
CHEN Lin;LI Jingjing;ZHOU Bo(State Key Laboratory of Cell Biology,Center for Excellence in Molecular Cell Science,Chinese Academy of Sciences,Shanghai 20003 J,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2021年第11期2169-2176,共8页
Chinese Journal of Cell Biology
基金
国家自然科学基金面上项目(批准号:81730006)资助的课题。
关键词
造血干细胞
流式细胞术
骨髓
hematopoietic stem cells
flow cytometry
bone marrow
