Abstract
Mesenchymal stem cells (MSCs) were initially characterized as connective tissue progenitors resident in bone marrow, but have now been isolated from a variety of tissues and organs and shown to also exhibit potent tissue regenerative properties mediated largely via paracrine actions. These findings have spurred the development of MSC-based therapies for treating a diverse array of nonskeletal diseases. Although genetic and experimental rodent models of disease represent important tools for developing efficacious MSC-based therapies, development of reliable methods to isolate MSCs from mouse bone marrow has been hampered by the unique biological properties of these cells. Indeed, few isolation schemes afford high yields and purity while maintaining the genomic integrity of cells. We recently demonstrated that mouse MSCs are highly sensitive to oxidative stress, and long-term expansion of these cells in atmospheric oxygen selects for immortalized clones that lack a functional p53 protein. Herein, we describe a protocol for the isolation of primary MSCs from mouse bone marrow that couples immunodepletion with culture in a low-oxygen environment and affords high purity and yield while preserving p53 function.
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Acknowledgments
The authors would like to thank previous members of the Phinney lab for assistance with development of this protocol including Dr. Gene C. Kopen, Maria Dutreil, and Melody Baddoo. This work is supported by a grant to DGP (1 R24-OD018254-01) from the Director’s Office at NIH, which provides funds to distribute IDmMSCs prepared from strains of interest to researchers worldwide. Contact dphinney@scripps.edu for more information.
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Boregowda, S.V., Krishnappa, V., Phinney, D.G. (2016). Isolation of Mouse Bone Marrow Mesenchymal Stem Cells. In: Gnecchi, M. (eds) Mesenchymal Stem Cells. Methods in Molecular Biology, vol 1416. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3584-0_11
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DOI: https://doi.org/10.1007/978-1-4939-3584-0_11
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