Murine hind limb long bone dissection and bone marrow isolation

Sarah R. Amend, Kenneth C. Valkenburg, Kenneth J. Pienta

Research output: Contribution to journalArticlepeer-review

Abstract

Investigation of the bone and the bone marrow is critical in many research fields including basic bone biology, immunology, hematology, cancer metastasis, biomechanics, and stem cell biology. Despite the importance of the bone in healthy and pathologic states, however, it is a largely under-researched organ due to lack of specialized knowledge of bone dissection and bone marrow isolation. Mice are a common model organism to study effects on bone and bone marrow, necessitating a standardized and efficient method for long bone dissection and bone marrow isolation for processing of large experimental cohorts. We describe a straightforward dissection procedure for the removal of the femur and tibia that is suitable for downstream applications, including but not limited to histomorphologic analysis and strength testing. In addition, we outline a rapid procedure for isolation of bone marrow from the long bones via centrifugation with limited handling time, ideal for cell sorting, primary cell culture, or DNA, RNA, and protein extraction. The protocol is streamlined for rapid processing of samples to limit experimental error, and is standardized to minimize user-to-user variability.

Original languageEnglish (US)
Article numbere53936
JournalJournal of Visualized Experiments
Volume2016
Issue number110
DOIs
StatePublished - Apr 2016

Keywords

  • Bone
  • Bone biology
  • Bone marrow
  • Cancer biology
  • Immunology
  • Immunology
  • Issue 110
  • Mouse dissection

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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