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Isolation of Mouse Bone Marrow Niche Cells for Single-Cell Sequencing
Yaphet Bustos1, Adrienne M Dorrance2, Chinmayee Goda3
1Division of Oncological Sciences, Huntsman Cancer Institute, University of Utah, Salt Lake City, UT, USA.
Methods in Molecular Biology (Clifton, N.J.)
|May 21, 2025
Summary
This chapter details a method to isolate bone marrow (BM) niche cells using fluorescence-activated cell sorting. This technique yields high-quality cells for single-cell RNA sequencing, advancing hematopoiesis research.
Area of Science:
- Hematology
- Cell Biology
- Genomics
Background:
- The bone marrow (BM) niche contains diverse cell types crucial for hematopoiesis and disease.
- Investigating these heterogeneous cell populations requires advanced techniques like single-cell RNA sequencing.
Purpose of the Study:
- To describe a method for isolating bone marrow niche cells from mouse femurs and tibias.
- To enable high-throughput analysis of cellular heterogeneity within the bone marrow microenvironment.
Main Methods:
- Isolation of bone marrow cells from femurs and tibias of mice.
- Purification of specific bone marrow niche cell populations using fluorescence-activated cell sorting (FACS).
- Preparation of isolated cells for downstream single-cell RNA sequencing (scRNA-seq).
Main Results:
- Successful isolation of high-quality bone marrow niche cells.
- Demonstration of high cell viability and purity post-isolation.
- Establishment of a reliable protocol for scRNA-seq applications.
Conclusions:
- The described FACS method provides a robust approach for isolating viable and pure bone marrow niche cells.
- This protocol facilitates detailed investigation of cellular composition and function in the BM niche.
- The method supports advancements in understanding hematopoiesis and related diseases through single-cell genomics.
