Advertisement

Isolation and Differentiation of Murine Primary Brown/Beige Preadipocytes

  • Hui Yu
  • Margo Emont
  • Heejin Jun
  • Jun WuEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1773)

Abstract

Studies on thermogenic adipose cells (brown and beige fat) hold promise to treat obesity and its associated metabolic disorders due to the ability of these cells to dissipate energy in the form of heat. Although immortalized adipose cell lines have been widely used to investigate cell autonomous regulations, many physiological functions of thermogenic fat need to be studied with primary fat cells. Here, we present a detailed protocol of the isolation, culture, and differentiation of primary brown and beige preadipocytes from the stromal vascular fraction of murine interscapular and inguinal adipose depots.

Key words

Beige fat cells Brown fat cells Stromal vascular fraction Preadipocytes 

References

  1. 1.
    Freedman DS (2011) Obesity - United States, 1988-2008. MMWR Surveill Summ 60(Suppl):73–77. su6001a15 [pii]Google Scholar
  2. 2.
    Ogden CL, Carroll MD, Kit BK, Flegal KM (2014) Prevalence of childhood and adult obesity in the United States, 2011-2012. JAMA 311(8):806–814. https://doi.org/10.1001/jama.2014.732CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Seale P, Bjork B, Yang W, Kajimura S, Chin S, Kuang S, Scime A, Devarakonda S, Conroe HM, Erdjument-Bromage H, Tempst P, Rudnicki MA, Beier DR, Spiegelman BM (2008) PRDM16 controls a brown fat/skeletal muscle switch. Nature 454(7207):961–967. https://doi.org/10.1038/nature07182. nature07182 [pii]CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Lepper C, Fan CM (2010) Inducible lineage tracing of Pax7-descendant cells reveals embryonic origin of adult satellite cells. Genesis 48(7):424–436. https://doi.org/10.1002/dvg.20630CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Wu J, Bostrom P, Sparks LM, Ye L, Choi JH, Giang AH, Khandekar M, Virtanen KA, Nuutila P, Schaart G, Huang K, Tu H, van Marken Lichtenbelt WD, Hoeks J, Enerback S, Schrauwen P, Spiegelman BM (2012) Beige adipocytes are a distinct type of thermogenic fat cell in mouse and human. Cell 150(2):366–376. https://doi.org/10.1016/j.cell.2012.05.016CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    van Marken Lichtenbelt WD, Vanhommerig JW, Smulders NM, Drossaerts JM, Kemerink GJ, Bouvy ND, Schrauwen P, Teule GJ (2009) Cold-activated brown adipose tissue in healthy men. N Engl J Med 360(15):1500–1508. https://doi.org/10.1056/NEJMoa0808718. 360/15/1500 [pii]CrossRefPubMedGoogle Scholar
  7. 7.
    Cypess AM, Lehman S, Williams G, Tal I, Rodman D, Goldfine AB, Kuo FC, Palmer EL, Tseng YH, Doria A, Kolodny GM, Kahn CR (2009) Identification and importance of brown adipose tissue in adult humans. N Engl J Med 360(15):1509–1517. https://doi.org/10.1056/NEJMoa0810780. 360/15/1509 [pii]CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Virtanen KA, Lidell ME, Orava J, Heglind M, Westergren R, Niemi T, Taittonen M, Laine J, Savisto NJ, Enerback S, Nuutila P (2009) Functional brown adipose tissue in healthy adults. N Engl J Med 360(15):1518–1525. https://doi.org/10.1056/NEJMoa0808949. 360/15/1518 [pii]CrossRefPubMedGoogle Scholar
  9. 9.
    Sharp LZ, Shinoda K, Ohno H, Scheel DW, Tomoda E, Ruiz L, Hu H, Wang L, Pavlova Z, Gilsanz V, Kajimura S (2012) Human BAT possesses molecular signatures that resemble beige/brite cells. PLoS One 7(11):e49452. https://doi.org/10.1371/journal.pone.0049452CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Lidell ME, Betz MJ, Dahlqvist Leinhard O, Heglind M, Elander L, Slawik M, Mussack T, Nilsson D, Romu T, Nuutila P, Virtanen KA, Beuschlein F, Persson A, Borga M, Enerback S (2013) Evidence for two types of brown adipose tissue in humans. Nat Med 19(5):631–634. https://doi.org/10.1038/nm.3017CrossRefPubMedGoogle Scholar
  11. 11.
    Jespersen NZ, Larsen TJ, Peijs L, Daugaard S, Homoe P, Loft A, de Jong J, Mathur N, Cannon B, Nedergaard J, Pedersen BK, Moller K, Scheele C (2013) A classical brown adipose tissue mRNA signature partly overlaps with brite in the supraclavicular region of adult humans. Cell Metab 17(5):798–805. https://doi.org/10.1016/j.cmet.2013.04.011CrossRefPubMedGoogle Scholar
  12. 12.
    Cypess AM, White AP, Vernochet C, Schulz TJ, Xue R, Sass CA, Huang TL, Roberts-Toler C, Weiner LS, Sze C, Chacko AT, Deschamps LN, Herder LM, Truchan N, Glasgow AL, Holman AR, Gavrila A, Hasselgren PO, Mori MA, Molla M, Tseng YH (2013) Anatomical localization, gene expression profiling and functional characterization of adult human neck brown fat. Nat Med 19(5):635–639. https://doi.org/10.1038/nm.3112CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Shinoda K, Luijten IH, Hasegawa Y, Hong H, Sonne SB, Kim M, Xue R, Chondronikola M, Cypess AM, Tseng YH, Nedergaard J, Sidossis LS, Kajimura S (2015) Genetic and functional characterization of clonally derived adult human brown adipocytes. Nat Med 21(4):389–394. https://doi.org/10.1038/nm.3819CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Yoneshiro T, Aita S, Matsushita M, Kayahara T, Kameya T, Kawai Y, Iwanaga T, Saito M (2013) Recruited brown adipose tissue as an antiobesity agent in humans. J Clin Invest 123(8):3404–3408. https://doi.org/10.1172/JCI67803CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    van der Lans AA, Hoeks J, Brans B, Vijgen GH, Visser MG, Vosselman MJ, Hansen J, Jorgensen JA, Wu J, Mottaghy FM, Schrauwen P, van Marken Lichtenbelt WD (2013) Cold acclimation recruits human brown fat and increases nonshivering thermogenesis. J Clin Invest 123(8):3395–3403. https://doi.org/10.1172/JCI68993CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Lee P, Smith S, Linderman J, Courville AB, Brychta RJ, Dieckmann W, Werner CD, Chen KY, Celi FS (2014) Temperature-acclimated brown adipose tissue modulates insulin sensitivity in humans. Diabetes 63(11):3686–3698. https://doi.org/10.2337/db14-0513CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Chondronikola M, Volpi E, Borsheim E, Porter C, Annamalai P, Enerback S, Lidell ME, Saraf MK, Labbe SM, Hurren NM, Yfanti C, Chao T, Andersen CR, Cesani F, Hawkins H, Sidossis LS (2014) Brown adipose tissue improves whole-body glucose homeostasis and insulin sensitivity in humans. Diabetes 63(12):4089–4099. https://doi.org/10.2337/db14-0746CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Green H, Kehinde O (1974) Sublines of mouse 3t3 cells that accumulate lipid. Cell 1(3):113–116. https://doi.org/10.1016/0092-8674(74)90126-3CrossRefGoogle Scholar
  19. 19.
    Green H, Kehinde O (1976) Spontaneous heritable changes leading to increased adipose conversion in 3T3 cells. Cell 7(1):105–113CrossRefPubMedGoogle Scholar
  20. 20.
    Ross SR, Choy L, Graves RA, Fox N, Solevjeva V, Klaus S, Ricquier D, Spiegelman BM (1992) Hibernoma formation in transgenic mice and isolation of a brown adipocyte cell line expressing the uncoupling protein gene. Proc Natl Acad Sci U S A 89(16):7561–7565CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Pond CM, Mattacks CA (2002) The activation of the adipose tissue associated with lymph nodes during the early stages of an immune response. Cytokine 17(3):131–139. https://doi.org/10.1006/cyto.2001.0999CrossRefPubMedGoogle Scholar
  22. 22.
    Benezech C, Mader E, Desanti G, Khan M, Nakamura K, White A, Ware CF, Anderson G, Caamano JH (2012) Lymphotoxin-beta receptor signaling through NF-kappaB2-RelB pathway reprograms adipocyte precursors as lymph node stromal cells. Immunity 37(4):721–734. https://doi.org/10.1016/j.immuni.2012.06.010CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Life Sciences InstituteUniversity of MichiganAnn ArborUSA
  2. 2.Department of Molecular and Integrative PhysiologyUniversity of MichiganAnn ArborUSA

Personalised recommendations