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Updated: Aug 16, 2026
Isolation, Culture, and Differentiation of Bone Marrow Stromal Cells and Osteoclast Progenitors from Mice
Published on: January 6, 2018
[Isolation and cultivation of mouse bone marrow mesenchymal stem cells]
Yi Zhang1, Liansan Zhao, Hong Tang
1West China Hospital of Sichuan University, Biotherapy of Human Diseases of Ministry of Education, Key Laboratory of Molecular Biology for Infectious Diseases of Sichuan Province, Chengdu 610041, China.
Abstract:
To establish the best condition of isolation and cultivation of mouse bone marrow mesenchymal stem cells (BMSCs) in vitro, we isolated BMSCs from BALB/C mouse using density gradient centrifugation and adherent selecting. The effects of different centrifuge power, adherent time, serum concentration and cell density on the isolation and cultivation of BMSCs were investigated. The best isolating condition is: 500g x 30min, 24 hours adhering. The best cell density is 12-20 x 10(5)/ml of primary cells and 6.4-25.6 x 10(4)/ml of secondary cells. The best serum concentration is 10%. More than 90% of subcultured cells were adhesive in 8 hours. Thus we have established a cell biological method of isolation and cultivation of BMSCs.
Insights
Researchers optimized conditions for isolating and cultivating mouse bone marrow mesenchymal stem cells (BMSCs). Key findings include optimal centrifugation, adherence time, serum concentration, and cell density for efficient BMSC culture.
Area of Science:
- Cell Biology
- Stem Cell Research
- Biotechnology
Background:
- Mouse bone marrow mesenchymal stem cells (BMSCs) are crucial for regenerative medicine.
- Optimizing in vitro isolation and cultivation is essential for their effective use.
Purpose of the Study:
- To determine the optimal conditions for isolating and cultivating mouse BMSCs.
- To establish a reliable cell biological method for BMSC propagation.
Main Methods:
- BMSCs were isolated from BALB/C mice using density gradient centrifugation and adherent selection.
- The study investigated the impact of varying centrifugation force, adherence time, serum concentration, and cell density.
Main Results:
- Optimal isolation involved 500g centrifugation for 30 minutes.
- Best cultivation conditions included 24 hours adherence, 10% serum concentration, and specific primary/secondary cell densities.
- Over 90% of subcultured cells showed adherence within 8 hours.
Conclusions:
- A refined protocol for isolating and cultivating mouse BMSCs has been established.
- The optimized method ensures efficient propagation and high viability of BMSCs for research applications.
