Neonatal Mouse Bone Marrow Isolation and Preparation of Bone Marrow-Derived Macrophages

Madhavi Annamanedi; Jordan K. Vance; Cory M. Robinson

doi:10.3791/66613

Rivista

Immunologia e infezioni

Phagocytic Assay Evaluation of Neonatal Bone Marrow Derived Macrophages

May 24, 2024

Author Spotlight: Developing a Reproducible Method for Isolating Bone Marrow-Derived Macrophages from Neonatal Mice to Advance Early Life Immune Responses

01:12 min

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Non-Enzymatic Method for Isolating Neonatal Mouse Bone Marrow Cells

01:36 min

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Differentiation of Neonatal Bone Marrow-Derived Macrophages Using a Supernatant of L929 Cells

01:59 min

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Assessing Phagocytic Function of Neonatal Bone Marrow-Derived Macrophages Using a Fluorescent Bacterial Uptake Assay

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Madhavi Annamanedi¹, Jordan K. Vance¹, Cory M. Robinson^1,2

¹Department of Microbiology, Immunology, & Cell Biology,West Virginia University School of Medicine, ²Vaccine Development Center,West Virginia University Health Sciences Center

Summary

This protocol describes a non-enzymatic and straightforward method for isolating 7-9-day-old neonatal mouse bone marrow cells and generating differentiated macrophages using a supernatant of L929 cells as a source of granulocyte colony-stimulating factor (M-CSF). The bone marrow-derived macrophages were further analyzed for surface antigens F4/80, CD206, CD11b, and functional competency.