Abstract
Hematopoietic stem cells (HSCs) are ultimately responsible for the lifelong renewal of all blood cell lineages. In the bone marrow (BM), HSCs reside in specialized microenvironments referred to as the “niche.” HSC niche consists of complex components including heterogeneous cell populations, growth factors, and extracellular matrix molecules. The crosstalk between HSCs and their niche is essential to regulate the survival, self-renewal, migration, quiescence, and differentiation of HSCs. The application of mice models with endogenous ablation of specific cell types, advanced imaging technologies, high-throughput single-cell RNA sequencing, and single-cell mass cytometry methods have provided deep insights into communications between HSCs and niche cells. In this chapter, we have focused on three important cell types in the BM niche: mesenchymal stem cells (MSCs), osteoblasts (OBs), and endothelial cells (ECs). In order to address the interaction between HSCs and these three cell populations in BM niche, we have described methodology for (1) collecting total BM from femur and tibia of C57BL/6 mice; (2) analyzing or sorting of MSCs, OBs, and ECs based on the selection of surface markers CD45, Ter119, CD31, Sca1, and CD51 with flow cytometry; and (3) co-culturing the sorted cells with purified HSCs for further functional assays of HSCs.
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Acknowledgments
The authors are supported by the National Heart, Lung, and Blood Institute of the National Institutes of Health under awards R01HL124015 (YL), R21HL140213 (YL), and R43AI45726 (YL) and the Markey Cancer Center’s Flow Cytometry and Immune Monitoring Core Shared Resource Facility (P30CA177558). We thank the Markey Cancer Center’s Research Communications Office for editing and graphics support.
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Zhao, X., Zhang, C., Cui, X., Liang, Y. (2020). Interactions of Hematopoietic Stem Cells with Bone Marrow Niche. In: Turksen, K. (eds) Stem Cell Renewal and Cell-Cell Communication. Methods in Molecular Biology, vol 2346. Humana, New York, NY. https://doi.org/10.1007/7651_2020_298
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DOI: https://doi.org/10.1007/7651_2020_298
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