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Isolation and Identification of Murine Bone Marrow-Derived Macrophages and Osteomacs from Neonatal and Adult Mice

  • Joydeep GhoshEmail author
  • Safa F. Mohamad
  • Edward F. Srour
Part of the Methods in Molecular Biology book series


Hematopoietic stem cells (HSCs) are regulated by multiple components of the hematopoietic niche, including bone marrow-derived macrophages and osteomacs. However, both macrophages and osteomacs are phenotypically similar. Thus, specific phenotypic markers are required to differentially identify the effects of osteomacs and bone marrow macrophages on different physiological processes, including hematopoiesis and bone remodeling. Here, we describe a protocol for isolation of murine bone marrow-derived macrophages and osteomacs from neonatal and adult mice and subsequent identification by multi-parametric flow cytometry using an 8-color antibody panel.


Bone digestion CD166 Hematopoietic niche Bone-marrow macrophages Multiparameter flow cytometry Osteomacs 



The authors thank Indiana University Melvin and Bren Simon Cancer Center Flow Cytometry Resource Facility (FCRF) for their outstanding technical help and support. FCRF is partially funded by National Cancer Institute grant P30 CA082709 and National Institute of Diabetes and Digestive and Kidney Diseases grant U54 DK106846. We also thank the support of the NIH instrumentation grant 1S10D012270 for partial funding of the FCRF.


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Copyright information

© Springer Science+Business Media New York 2018

Authors and Affiliations

  • Joydeep Ghosh
    • 1
    Email author
  • Safa F. Mohamad
    • 2
  • Edward F. Srour
    • 1
    • 3
  1. 1.Department of MedicineIndiana University School of MedicineIndianapolisUSA
  2. 2.Department of Microbiology and ImmunologyIndiana University School of MedicineIndianapolisUSA
  3. 3.Department of Microbiology and ImmunologyIndiana University School of MedicineIndianapolisUSA

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