Skip to main content

Flow Cytometry Assisted Isolation of Adipose Tissue Derived Stem Cells

  • Protocol
  • First Online:
Book cover Thermogenic Fat

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1566))

Abstract

Adipose tissue dysfunction is typically seen in metabolic diseases, particularly obesity and diabetes. White adipocytes store fat while brown adipocyte dissipates it via thermogenesis. In addition, beige adipocytes develop in white fat depots in response to stimulation of β-adrenergic pathways. It appears that the three types of adipocytes—white, brown, and beige—can be formed de novo from stem/precursor cells or via transdifferentiation. Identifying the presumptive progenitors that harbor capacity to differentiate to these distinct adipocyte cell types will enable their functional characterization. Moreover, the presence or absence of white/brown/beige adipocytes is correlated with metabolic dysfunction making their study of medical relevance. Robust, reliable, and reproducible methods of identification and isolation of adipocyte progenitors will stimulate further detailed understanding of white, brown, and beige adipogenesis.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 119.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 159.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Sun K, Kusminski CM, Scherer PE (2011) Adipose tissue remodeling and obesity. J Clin Invest 121(6):2094–2101. doi:10.1172/JCI45887

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Gesta S, Tseng YH, Kahn CR (2007) Developmental origin of fat: tracking obesity to its source. Cell 131(2):242–256. doi:10.1016/j.cell.2007.10.004

    Article  CAS  PubMed  Google Scholar 

  3. Rosen ED, Spiegelman BM (2014) What we talk about when we talk about fat. Cell 156(1-2):20–44. doi:10.1016/j.cell.2013.12.012

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Cannon B, Nedergaard J (2004) Brown adipose tissue: function and physiological significance. Physiol Rev 84(1):277–359. doi:10.1152/physrev.00015.2003

    Article  CAS  PubMed  Google Scholar 

  5. Nedergaard J, Bengtsson T, Cannon B (2007) Unexpected evidence for active brown adipose tissue in adult humans. Am J Physiol Endocrinol Metab 293(2):E444–E452. doi:10.1152/ajpendo.00691.2006

    Article  CAS  PubMed  Google Scholar 

  6. 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. doi:10.1056/NEJMoa0810780

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Nedergaard J, Cannon B (2010) The changed metabolic world with human brown adipose tissue: therapeutic visions. Cell Metab 11(4):268–272. doi:10.1016/j.cmet.2010.03.007

    Article  CAS  PubMed  Google Scholar 

  8. Enerback S (2010) Human brown adipose tissue. Cell Metab 11 (4):248–252.doi:10.1016/j.cmet.2010.03.008, S1550-4131(10)00078-1 [pii]

  9. Cousin B, Cinti S, Morroni M, Raimbault S, Ricquier D, Penicaud L, Casteilla L (1992) Occurrence of brown adipocytes in rat white adipose tissue: molecular and morphological characterization. J Cell Sci 103(Pt 4):931–942

    CAS  PubMed  Google Scholar 

  10. Guerra C, Koza RA, Yamashita H, Walsh K, Kozak LP (1998) Emergence of brown adipocytes in white fat in mice is under genetic control. Effects on body weight and adiposity. J Clin Invest 102(2):412–420. doi:10.1172/JCI3155

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Xue B, Rim JS, Hogan JC, Coulter AA, Koza RA, Kozak LP (2007) Genetic variability affects the development of brown adipocytes in white fat but not in interscapular brown fat. J Lipid Res 48(1):41–51

    Article  CAS  PubMed  Google Scholar 

  12. Petrovic N, Walden TB, Shabalina IG, Timmons JA, Cannon B, Nedergaard J (2010) Chronic peroxisome proliferator-activated receptor gamma (PPAR gamma) activation of epididymally derived white adipocyte cultures reveals a population of thermogenically competent, UCP1-containing adipocytes molecularly distinct from classic brown adipocytes. J Biol Chem 285(10):7153–7164

    Article  CAS  PubMed  Google Scholar 

  13. 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. doi:10.1016/j.cell.2012.05.016

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Schulz TJ, Huang TL, Tran TT, Zhang H, Townsend KL, Shadrach JL, Cerletti M, McDougall LE, Giorgadze N, Tchkonia T, Schrier D, Falb D, Kirkland JL, Wagers AJ, Tseng YH (2011) Identification of inducible brown adipocyte progenitors residing in skeletal muscle and white fat. Proc Natl Acad Sci U S A 108(1):143–148. doi:10.1073/pnas.1010929108

    Article  CAS  PubMed  Google Scholar 

  15. Wang QA, Tao C, Gupta RK, Scherer PE (2013) Tracking adipogenesis during white adipose tissue development, expansion and regeneration. Nat Med 19(10):1338–1344. doi:10.1038/nm.3324

    Article  PubMed  PubMed Central  Google Scholar 

  16. Himms-Hagen J, Melnyk A, Zingaretti MC, Ceresi E, Barbatelli G, Cinti S (2000) Multilocular fat cells in WAT of CL-316243-treated rats derive directly from white adipocytes. Am J Physiol 279(3):C670–C681

    CAS  Google Scholar 

  17. Lee YH, Petkova AP, Mottillo EP, Granneman JG (2012) In vivo identification of bipotential adipocyte progenitors recruited by beta3-adrenoceptor activation and high-fat feeding. Cell Metab 15(4):480–491. doi:10.1016/j.cmet.2012.03.009

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Bartelt A, Heeren J (2014) Adipose tissue browning and metabolic health. Nat Rev Endocrinol 10(1):24–36. doi:10.1038/nrendo.2013.204

    Article  CAS  PubMed  Google Scholar 

  19. Harms M, Seale P (2013) Brown and beige fat: development, function and therapeutic potential. Nat Med 19(10):1252–1263. doi:10.1038/nm.3361

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Sushil G. Rane .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Wankhade, U.D., Rane, S.G. (2017). Flow Cytometry Assisted Isolation of Adipose Tissue Derived Stem Cells. In: Wu, J. (eds) Thermogenic Fat. Methods in Molecular Biology, vol 1566. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6820-6_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-6820-6_3

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6819-0

  • Online ISBN: 978-1-4939-6820-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics