Abstract
Murine models of obesity or reduced adiposity are a valuable resource for understanding the role of adipocyte dysfunction in metabolic disorders. Adipose tissue stromal vascular cells or primary adipocytes derived from murine adipose tissue and grown in culture are essential tools for studying the mechanisms underlying adipocyte development and function. Herein, we describe methods for the isolation, expansion, and long-term storage of murine adipose-derived stromal/stem cells along with protocols for inducing adipogenesis in this cell population or isolating the adipose stromal vascular fraction cells for flow cytometric analysis.
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References
Cawthorn WP, Scheller EL, MacDougald OA (2012) Adipose tissue stem cells meet preadipocyte commitment: going back to the future. J Lipid Res 53(2):227–246. https://doi.org/10.1194/jlr.R021089
Bourin P, Bunnell BA, Casteilla L, Dominici M, Katz AJ, March KL, Redl H, Rubin JP, Yoshimura K, Gimble JM (2013) Stromal cells from the adipose tissue-derived stromal vascular fraction and culture expanded adipose tissue-derived stromal/stem cells: a joint statement of the International Federation for Adipose Therapeutics and Science (IFATS) and the International Society for Cellular Therapy (ISCT). Cytotherapy 15(6):641–648. https://doi.org/10.1016/j.jcyt.2013.02.006
Kelly KA, Tanaka S, Baron R, Gimble JM (1998) Murine bone marrow stromally derived BMS2 adipocytes support differentiation and function of osteoclast-like cells in vitro. Endocrinology 139(4):2092–2101
Thompson DL, Lum KD, Nygaard SC, Kuestner RE, Kelly KA, Gimble JM, Moore EE (1998) The derivation and characterization of stromal cell lines from the bone marrow of p53−/− mice: new insights into osteoblast and adipocyte differentiation. J Bone Miner Res 13(2):195–204
Safford KM, Hicok KC, Safford SD, Halvorsen YD, Wilkison WO, Gimble JM, Rice HE (2002) Neurogenic differentiation of murine and human adipose-derived stromal cells. Biochem Biophys Res Commun 294(2):371–379
Zheng B, Cao B, Li G, Huard J (2006) Mouse adipose-derived stem cells undergo multilineage differentiation in vitro but primarily osteogenic and chondrogenic differentiation in vivo. Tissue Eng 12(7):1891–1901. https://doi.org/10.1089/ten.2006.12.1891
Grant RW, Dixit VD (2015) Adipose tissue as an immunological organ. Obesity 23(3):512–518. https://doi.org/10.1002/oby.21003
Grant R, Youm YH, Ravussin A, Dixit VD (2013) Quantification of adipose tissue leukocytosis in obesity. Methods Mol Biol 1040:195–209. https://doi.org/10.1007/978-1-62703-523-1_15
Bluher M, Patti ME, Gesta S, Kahn BB, Kahn CR (2004) Intrinsic heterogeneity in adipose tissue of fat-specific insulin receptor knock-out mice is associated with differences in patterns of gene expression. J Biol Chem 279(30):31891–31901. https://doi.org/10.1074/jbc.M404569200. M404569200 [pii]
Acknowledgment
This work is supported by the National Institutes of Health (NIDDK, 1R01DK099625, Z.E.F.). This work used the Cell Biology and Bioimaging Core facility at Pennington Biomedical Research Center that is supported in part by COBRE (NIH 8P20-GM103528) and NORC (NIH 2P30-DK072476) center grants from the National Institutes of Health.
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Kilroy, G., Dietrich, M., Wu, X., Gimble, J.M., Floyd, Z.E. (2018). Isolation of Murine Adipose-Derived Stromal/Stem Cells for Adipogenic Differentiation or Flow Cytometry-Based Analysis. In: Bunnell, B.A., Gimble, J.M. (eds) Adipose-Derived Stem Cells. Methods in Molecular Biology, vol 1773. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7799-4_11
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DOI: https://doi.org/10.1007/978-1-4939-7799-4_11
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7797-0
Online ISBN: 978-1-4939-7799-4
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