Abstract
Appropriate cell models are necessary for the investigation of thermogenic beige adipocyte differentiation from progenitor cells. Here, we describe a primary cell culture method that is based on defined progenitor cells from murine white adipose tissue and aims at minimizing confounding factors including cell heterogeneity and nonphysiological differentiation inducers. Adipocyte progenitor cells are enriched by immuno-magnetic separation, expanded minimally, and induced for beige adipocyte differentiation with carbaprostacyclin, a stable analogue of the endogenous mediator PGI2.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Diaz MB, Herzig S, Vegiopoulos A (2014) Thermogenic adipocytes: from cells to physiology and medicine. Metabolism 63(10):1238–1249. doi:10.1016/j.metabol.2014.07.002
Kajimura S, Spiegelman BM, Seale P (2015) Brown and Beige fat: physiological roles beyond heat generation. Cell Metab 22(4):546–559. doi:10.1016/j.cmet.2015.09.007
Sidossis L, Kajimura S (2015) Brown and beige fat in humans: thermogenic adipocytes that control energy and glucose homeostasis. J Clin Invest 125(2):478–486. doi:10.1172/JCI78362
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 USA 108(1):143–148. doi:10.1073/pnas.1010929108
Petrovic N, Walden TB, Shabalina IG, Timmons JA, Cannon B, Nedergaard J (2010) Chronic peroxisome proliferator-activated receptor gamma (PPARgamma) 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. doi:10.1074/jbc.M109.053942
Lehmann JM, Moore LB, Smitholiver TA, Wilkison WO, Willson TM, Kliewer SA (1995) An antidiabetic thiazolidinedione is a high-affinity ligand for peroxisome proliferator-activated receptor gamma(Ppar-Gamma). J Biol Chem 270(22):12953–12956
Samarasinghe SP, Sutanto MM, Danos AM, Johnson DN, Brady MJ, Cohen RN (2009) Altering PPARgamma ligand selectivity impairs adipogenesis by thiazolidinediones but not hormonal inducers. Obesity (Silver Spring) 17(5):965–972. doi:10.1038/oby.2008.629
Bayindir I, Babaeikelishomi R, Kocanova S, Sousa IS, Lerch S, Hardt O, Wild S, Bosio A, Bystricky K, Herzig S, Vegiopoulos A (2015) Transcriptional pathways in cPGI2-induced adipocyte progenitor activation for browning. Front Endocrinol(Lausanne) 6:129. doi:10.3389/fendo.2015.00129
Rodeheffer MS, Birsoy K, Friedman JM (2008) Identification of white adipocyte progenitor cells in vivo. Cell 135(2):240–249. doi:10.1016/j.cell.2008.09.036
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
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
Vishvanath L, MacPherson KA, Hepler C, Wang QA, Shao M, Spurgin SB, Wang MY, Kusminski CM, Morley TS, Gupta RK (2015) Pdgfrbeta mural preadipocytes contribute to adipocyte hyperplasia induced by high-fat-diet feeding and prolonged cold exposure in adult mice. Cell Metab. doi:10.1016/j.cmet.2015.10.018
Madsen L, Pedersen LM, Lillefosse HH, Fjaere E, Bronstad I, Hao Q, Petersen RK, Hallenborg P, Ma T, De Matteis R, Araujo P, Mercader J, Bonet ML, Hansen JB, Cannon B, Nedergaard J, Wang J, Cinti S, Voshol P, Doskeland SO, Kristiansen K (2010) UCP1 induction during recruitment of brown adipocytes in white adipose tissue is dependent on cyclooxygenase activity. PLoS One 5(6):e11391. doi:10.1371/journal.pone.0011391
Vegiopoulos A, Muller-Decker K, Strzoda D, Schmitt I, Chichelnitskiy E, Ostertag A, Berriel Diaz M, Rozman J, Hrabe de Angelis M, Nusing RM, Meyer CW, Wahli W, Klingenspor M, Herzig S (2010) Cyclooxygenase-2 controls energy homeostasis in mice by de novo recruitment of brown adipocytes. Science 328(5982):1158–1161. doi:10.1126/science.1186034
Brun RP, Tontonoz P, Forman BM, Ellis R, Chen J, Evans RM, Spiegelman BM (1996) Differential activation of adipogenesis by multiple PPAR isoforms. Genes Dev 10(8):974–984
Narumiya S, Sugimoto Y, Ushikubi F (1999) Prostanoid receptors: structures, properties, and functions. Physiol Rev 79(4):1193–1226
Ghandour RA, Giroud M, Vegiopoulos A, Herzig S, Ailhaud G, Amri EZ, Pisani DF (2016) IP-receptor and PPARs trigger the conversion of human white to brite adipocyte induced by carbaprostacyclin. Biochim Biophys Acta. doi:10.1016/j.bbalip.2016.01.007
Acknowledgments
This work was supported by the Deutsche Forschungsgemeinschaft (HE 3260/8-1), the Helmholtz Association (“Metabolic Dysfunction”), and the Human Frontier Science Program (RGY0082/2014).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Babaei, R., Bayindir-Buchhalter, I., Meln, I., Vegiopoulos, A. (2017). Immuno-Magnetic Isolation and Thermogenic Differentiation of White Adipose Tissue Progenitor 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_5
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6820-6_5
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