Isolation of Murine Neonatal and Adult Osteomacs to Examine Their Role in the Hematopoietic Niche

Safa F Mohamad; Melissa A Kacena

doi:10.1007/7651_2024_535

Isolation of Murine Neonatal and Adult Osteomacs to Examine Their Role in the Hematopoietic Niche

Methods Mol Biol. 2024 Mar 21. doi: 10.1007/7651_2024_535. Online ahead of print.

Authors

Safa F Mohamad¹, Melissa A Kacena²

Affiliations

¹ Department of Hematology and Oncology, Boston Children's Hospital/Harvard School of Medicine, Boston, MA, USA. Safa.Mohamad@childrens.harvard.edu.
² Department of Orthopaedic Surgery, Indiana University School of Medicine, Indianapolis, IN, USA.

PMID: 38507212
DOI: 10.1007/7651_2024_535

Abstract

Maintenance of hematopoietic stem cell (HSC) function is an orchestrated event between multiple cell types, and crosstalk between these cell types is an essential part of HSC regulation. Among the cell groups of the niche involved in this process are a group of bone-resident macrophages known as osteomacs (OM). Previously, it was demonstrated that OM and osteoblasts contained within neonatal calvarial cells are critical to maintain hematopoietic function. Additionally, interactions between neonatal calvarial cells and megakaryocytes further enhance this hematopoietic activity. In this chapter, we explore one such interaction involving OM and osteoblasts in the hematopoietic niche. We describe a protocol to isolate OM from both neonatal and adult mice, and subsequently use colony-forming assays to demonstrate their interaction with osteoblasts in maintaining HSC function.

Keywords: Colony-forming assays; Hematopoietic niche; Macrophages; Osteoblasts; Osteomacs.