Flow Cytometric Analysis of Mouse Hematopoietic Stem Cells
CHEN Lin, LI Jingjing, ZHOU Bo*
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-catulinGFP 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.
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