Murine Hind Limb Long Bone Dissection and Bone Marrow Isolation

作者：Amend Sarah R^*; Valkenburg Kenneth C; Pienta Kenneth J

来源：Jove-Journal of Visualized Experiments, 2016, (110): e53936.

DOI：10.3791/53936

• Immunology
• Issue 110
• Bone
• bone marrow
• mouse dissection
• bone biology
• cancer biology
• immunology

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.

• 出版日期2016-4