Abstract
The bone marrow (BM) of numerous species, including rodents and man, contains a rare population of cells termed marrow stromal cells or mesenchymal stem cells (MSC). Given the ability of these cells to differentiate into cells of the osteogenic, chondrogenic and adipogenic lineages, there is considerable interest in utilizing MSCs in a broad repertoire of cell-based therapies for the treatment of human disease. Before such potential therapies can be realized, a preclinical animal model in which to test and refine strategies utilizing MSC is required. Here we describe methods for the isolation of a highly enriched population of MSC from mouse cortical/compact bone (CB), quantitation using the colony forming unit-fibroblast assay (CFU-F) and in vitro expansion. These cells are both multipotent and capable of extensive in vitro expansion and thus represent an ideal cellular source to explore both the biological properties of MSC as well as their potential efficacy in a variety of cellular therapies.
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References
Simmons PJ, Torok-Storb B (1991) Identification of stromal cell precursors inhuman bone marrow by a novel monoclonal antibody, STRO-1. Blood 78(1):55–62
Gronthos S et al (2003) Molecular and cellular characterisation of highly purified stromal stem cells derived from human bone marrow. J Cell Sci 116(Pt 9):1827–1835
Phinney DG et al (1999) Plastic adherent stromal cells from the bone marrow of commonly used strains of inbred mice: variations in yield, growth, and differentiation. J Cell Biochem 72(4):570–585
Friedenstein AJ, Gorskaja JF, Kulagina NN (1976) Fibroblast precursors in normal and irradiated mouse hematopoietic organs. Exp Hematol 4(5):267–274
Taichman RS, Wang Z, Shiozawa Y, Jung Y, Song J, Balduino A, Wang J, Patel LR, Havens AM, Kucia M, Ratajczak MZ, Krebsbach PH (2010) Prospective identification and skeletal localization of cells capable of multilineage differentiation in vivo. Stem Cells Dev 19(10):1557–1570
Morikawa S, Mabuchi Y, Kubota Y, Nagai Y, Niibe K, Hiratsu E, Suzuki S, Miyauchi-Hara C, Nagoshi N, Sunabori T, Shimmura S, Miyawaki A, Nagagawa T, Suda T, Okano H, Matsuzaki Y (2009) Prospective identification, isolation, and systemic transplantation of multipotent mesenchymal stem cells in murine bone marrow. J Exp Med 206(11):2483–2496
Zhu H, Guo ZK, Jiang XX, Li H, Wang XY, Yao HY, Zhang Y, Mao N (2010) A protocol for isolation and culture of mesenchymal stem cells from mouse compact bone. Nat Protoc 5:550–560
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Short, B., Wagey, R. (2013). Isolation and Culture of Mesenchymal Stem Cells from Mouse Compact Bone. In: Helgason, C., Miller, C. (eds) Basic Cell Culture Protocols. Methods in Molecular Biology, vol 946. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-128-8_21
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DOI: https://doi.org/10.1007/978-1-62703-128-8_21
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-127-1
Online ISBN: 978-1-62703-128-8
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