Isolation and Culture of Bone Marrow Stromal Cells from a Mouse Tibia

Abstract

Source: Maridas, D. E., et al. Isolation, Culture, and Differentiation of Bone Marrow Stromal Cells and Osteoclast Progenitors from Mice. J. Vis. Exp.(2018)

In this video, we demonstrate the isolation and culture of bone marrow stromal cells from the long bones of mice. The isolated adherent BMSCs can be used for further analysis.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Harvest of Bones from Mice

1. Euthanize the animals using CO₂ followed by cervical dislocation and spray them with 70% ethanol.
   NOTE: Ensure that animals of the same sex and preferably the same age are used. We routinely use animals aged 7 to 8 weeks. We recommend pooling cells from at least 3 animals per experimental group to yield a better representative and reproducible cell population.
2. Place the euthanized mouse on its ba....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
200μL pipet tips	Rainin	17014401	Plasticware
1.5mL centrifuge tubes	USA Scientific	1615-5500	Plasticware
15mL conical tube	VWR	89039-668	Plasticware
50mL conical tube	VWR	89039-660	Plasticware
Phosphate-Buffered Saline (PBS)	Sigma-Aldrich	21-040-CM	Reagent
Ethanol	Fisher Science	04-355-451	Reagent
Dissection tools
70μm filters	BD Falcon	352350
0.25% trypsin	Gibco	25200	Reagent
Kimwipe	VWR	82003-820