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Updated: Feb 8, 2026

Application of Retinoic Acid to Obtain Osteocytes Cultures from Primary Mouse Osteoblasts
Published on: May 13, 2014
Osteoblast Sorting and Intracellular Staining of CXCL12
Weihuan Wang1, Gurnoor Majihail2, Cui Lui1
1Department of Pathology, Case Western Reserve University, Cleveland, OH, USA.
Abstract:
Osteoblasts are bone marrow endosteum-lining niche cells playing important roles in the regulation of hematopoietic stem cells by secreting factors and cell adhesion molecules. Characterization of primary osteoblasts has been achieved through culture of outgrowth of collagenase treated bone. Immunophenotyping and flow-based analysis of long bone osteoblasts offer a simplified and rapid approach to characterize osteoblasts. We describe a modified procedure of isolating mouse bone marrow osteoblastic cells based on cell surface immunophenotyping. The chemokine CXCL12 (also known as stromal-derived factor, SDF-1) together with its receptor CXCR4 are expressed by osteoblasts and bone marrow stroma cells. The CXCL12-CXCR4 axis is important for hematopoietic stem cell retention to their niches (Sugiyama et al., 2006) and for supporting leukemia initiating cell activity (Pitt et al., 2015). Here we describe the procedure of intracellular staining of CXCL12.
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