Home › Immunology › Murine Hind Limb Long Bone Dissection and Bone Marrow Isolation
Immunology JoVE (Open Access) Citable · DOI

Murine Hind Limb Long Bone Dissection and Bone Marrow Isolation

DOI: 10.3791/53936-v
What you'll learn
  • ✓ Dissect mouse femurs and tibiae using sterile technique
  • ✓ Prepare long bones for bone marrow extraction
  • ✓ Isolate bone marrow via centrifugation-based method
  • ✓ Apply harvested bone marrow to downstream analyses
Protocol

Biopharma Insights Here we present a protocol for the dissection of hind limb long bones (femurs and tibiae) from the laboratory mouse. We further describe a rapid technique for bone marrow isolation from these bones that utilizes centrifugation for removal of bone marrow from the bone marrow space.

Difficulty
intermediate
Total time
~15-20 min per mouse (dissection and isolation)
Model organism
Mouse (laboratory, strain not specified)
Biosafety
BSL-1

Steps

1
Dissect hind limb long bones from mouse

Remove femurs and tibiae from euthanized mouse hind limbs using aseptic surgical technique. Clean soft tissue from bone surfaces.

▶ 00:41
2
Prepare long bones for marrow isolation

Trim and section dissected femurs and tibiae into appropriate fragments to facilitate bone marrow access and centrifugation.

▶ 03:01
3
Isolate bone marrow via centrifugation

Place prepared bone fragments in tubes and centrifuge at specified parameters to extract bone marrow from the marrow cavity.

▶ 04:25
4
Apply isolated marrow to downstream analyses

Demonstrate representative downstream applications of harvested bone marrow and dissected long bones for research purposes.

▶ 05:29
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