Abstract
Perivascular adipose tissue (PVAT) has the capacity to secrete vasoactive mediators with the potential to regulate vascular function. Given its location adjacent to the vasculature, PVAT dysfunction may be part of the pathophysiology of cardiovascular diseases. To study the mechanisms of PVAT dysfunction, several adipogenic models have been proposed. However, these approaches do not adequately reflect PVAT adipocyte phenotypes variability that depends on their anatomical location. Despite PVAT importance in modulating vascular function, to date, there is not a depot-specific adipogenic model for PVAT adipocytes. We present a model that uses coculturing of PVAT stromal vascular fraction derived preadipocytes with primary adipocytes isolated from the same PVAT. Preadipocytes were isolated from thoracic aorta PVAT and mesenteric resistance artery PVAT (mPVAT). Upon confluency, cells were induced to differentiate for 7 and 14 days using a standard protocol (SP) or standard protocol cocultured with primary adipocytes isolated from the same adipose depots (SPA) for 96, 120, and 144 h. SPA reduced the time for differentiation of stromal vascular fraction derived preadipocytes and increased their capacity to store lipids compared with SP as indicated by lipid accumulation, lipolytic responses, gene marker profile expression, and adiponectin secretion. The coculture system improved adipogenesis efficiency by enhancing lipid accumulation and reducing the time of induction, therefore, is a more efficient method compared to SP alone.
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Baglioni S, Cantini G, Poli G, Francalanci M, Squecco R, Di Franco A, Borgogni E, Frontera S, Nesi G, Liotta F, Lucchese M, Perigli G, Francini F, Forti G, Serio M, Luconi M (2012) Functional differences in visceral and subcutaneous fat pads originate from differences in the adipose stem cell. PLoS ONE 7:e36569
Brzoska M, Geiger H, Gauer S, Baer P (2005) Epithelial differentiation of human adipose tissue-derived adult stem cells. Biochem Biophys Res Commun 330:142–150
Chatterjee TK, Stoll LL, Denning GM, Harrelson A, Blomkalns AL, Idelman G, Rothenberg FG, Neltner B, Romig-Martin SA, Dickson EW, Rudich S, Weintraub NL (2009) Proinflammatory phenotype of perivascular adipocytes: influence of high-fat feeding. Circ Res 104:541–549
Contreras GA, Thelen K, Ayala-Lopez N, Watts SW (2016) The distribution and adipogenic potential of perivascular adipose tissue adipocyte progenitors is dependent on sexual dimorphism and vessel location. Physiol Rep. 4:e12993
Contreras GA, Strieder-Barboza C, de Souza J, Gandy J, Mavangira V, Lock AL, Sordillo LM (2017) Periparturient lipolysis and oxylipid biosynthesis in bovine adipose tissues. PLoS ONE 12:e0188621
Ebke LA, Nestor-Kalinoski AL, Slotterbeck BD, Al-Dieri AG, Ghosh-Lester S, Russo L, Najjar SM, von Grafenstein H, McInerney MF (2014) Tight association between macrophages and adipocytes in obesity: implications for adipocyte preparation. Obesity (Silver Spring, Md.) 22:1246–1255
Fried SK, Moustaid-Moussa N (2001) Culture of Adipose Tissue and Isolated Adipocytes. In: Ailhaud G (ed) Adipose tissue protocols. Springer, New York, pp 197–212
Fuster JJ, Ouchi N, Gokce N, Walsh K (2016) Obesity-induced changes in adipose tissue microenvironment and their impact on cardiovascular disease. Circ Res 118:1786–1807
Ismail A, Ayala-Lopez N, Ahmad M, Watts SW (2017) 3T3-L1 cells and perivascular adipocytes are not equivalent in amine transporter expression. FEBS Lett 591:137–144
Knebel B, Goeddeke S, Poschmann G, Markgraf DF, Jacob S, Nitzgen U, Passlack W, Preuss C, Dicken HD, Stuhler K, Hartwig S, Lehr S, Kotzka J (2017) Novel insights into the adipokinome of obese and obese/diabetic mouse models. Int J Mol Sci 18:E1928. https://doi.org/10.3390/ijms18091928
Kokta TA, Strat AL, Papasani MR, Szasz JI, Dodson MV, Hill RA (2008) Regulation of lipid accumulation in 3T3-L1 cells: insulin-independent and combined effects of fatty acids and insulin. Animal Int J Animal Biosci 2:92–99
Kusuyama J, Komorizono A, Bandow K, Ohnishi T, Matsuguchi T (2016) CXCL3 positively regulates adipogenic differentiation. J Lipid Res 57:1806–1820
Lian X, Gollasch M (2016) A clinical perspective: contribution of dysfunctional perivascular adipose tissue (PVAT) to cardiovascular risk. Curr Hypertens Rep 18:82
Lihn AS, Bruun JM, He G, Pedersen SB, Jensen PF, Richelsen B (2004) Lower expression of adiponectin mRNA in visceral adipose tissue in lean and obese subjects. Mol Cell Endocrinol 219:9–15
Macotela Y, Emanuelli B, Mori MA, Gesta S, Schulz TJ, Tseng YH, Kahn CR (2012) Intrinsic differences in adipocyte precursor cells from different white fat depots. Diabetes 61:1691–1699
Marshall S, Garvey WT, Geller M (1984) Primary culture of isolated adipocytes. A new model to study insulin receptor regulation and insulin action. J Biol Chem 259:6376–6384
Mitchell JB, McIntosh K, Zvonic S, Garrett S, Floyd ZE, Kloster A, Di Halvorsen Y, Storms RW, Goh B, Kilroy G, Wu X, Gimble JM (2006) Immunophenotype of human adipose-derived cells: temporal changes in stromal-associated and stem cell-associated markers. Stem cells (Dayton, Ohio). 24:376–385
Morimoto C, Kameda K, Tsujita T, Okuda H (2001) Relationships between lipolysis induced by various lipolytic agents and hormone-sensitive lipase in rat fat cells. J Lipid Res 42:120–127
Moseti D, Regassa A, Kim WK (2016) Molecular regulation of adipogenesis and potential anti-adipogenic bioactive molecules. Int J Mol Sci 17:E124
Ouwens DM, Sell H, Greulich S, Eckel J (2010) The role of epicardial and perivascular adipose tissue in the pathophysiology of cardiovascular disease. J Cell Mol Med 14:2223–2234
Padilla J, Jenkins NT, Vieira-Potter VJ, Laughlin MH (2013) Divergent phenotype of rat thoracic and abdominal perivascular adipose tissues. Am J Physiol Regul, Integr Comp Physiol 304:R543–R552
Payne GA, Bohlen HG, Dincer UD, Borbouse L, Tune JD (2009) Periadventitial adipose tissue impairs coronary endothelial function via PKC-beta-dependent phosphorylation of nitric oxide synthase. Am J Physiol Heart Circ Physiol 297:H460–H465
Rajsheker S, Manka D, Blomkalns AL, Chatterjee TK, Stoll LL, Weintraub NL (2010) Crosstalk between perivascular adipose tissue and blood vessels. Curr Opin Pharmacol 10:191–196
Roca-Rivada A, Alonso J, Al-Massadi O, Castelao C, Peinado JR, Seoane LM, Casanueva FF, Pardo M (2011) Secretome analysis of rat adipose tissues shows location-specific roles for each depot type. J Proteom 74:1068–1079
Ruiz-Ojeda FJ, Rupérez AI, Gomez-Llorente C, Gil A, Aguilera CM (2016) Cell models and their application for studying adipogenic differentiation in relation to obesity: a review. Int J Mol Sci 17:E1040
Sanchez-Gurmaches J, Hung CM, Guertin DA (2016) Emerging complexities in adipocyte origins and identity. Trends Cell Biol 26:313–326
Shoham N, Gefen A (2011) Stochastic modeling of adipogenesis in 3T3-L1 cultures to determine probabilities of events in the cell’s life cycle. Ann Biomed Eng 39:2637–2653
Stacey DH, Hanson SE, Lahvis G, Gutowski KA, Masters KS (2009) In vitro adipogenic differentiation of preadipocytes varies with differentiation stimulus, culture dimensionality, and scaffold composition. Tissue Eng Part A 15:3389–3399
Tchkonia T, Giorgadze N, Pirtskhalava T, Tchoukalova Y, Karagiannides I, Forse RA, DePonte M, Stevenson M, Guo W, Han J, Waloga G, Lash TL, Jensen MD, Kirkland JL (2002) Fat depot origin affects adipogenesis in primary cultured and cloned human preadipocytes. Am J Physiol Regul Integr Comp Physiol 282:R1286–R1296
Tchkonia T, Giorgadze N, Pirtskhalava T, Thomou T, DePonte M, Koo A, Forse RA, Chinnappan D, Martin-Ruiz C, von Zglinicki T, Kirkland JL (2006) Fat depot-specific characteristics are retained in strains derived from single human preadipocytes. Diabetes 55:2571–2578
Thelen K, Ayala-Lopez N, Watts SW, Contreras GA (2017) Expansion and adipogenesis induction of adipocyte progenitors from perivascular adipose tissue isolated by magnetic activated cell sorting. J Vis Exp. https://doi.org/10.3791/55818
Tran KV, Fitzgibbons T, Min SY, DeSouza T, Corvera S (2018) Distinct adipocyte progenitor cells are associated with regional phenotypes of perivascular aortic fat in mice. Mol Metab 9:199–206
Vargas D, Camacho J, Duque J, Carreno M, Acero E, Perez M, Ramirez S, Umana J, Obando C, Guerrero A, Sandoval N, Rodriguez G, Lizcano F (2017) Functional characterization of preadipocytes derived from human periaortic adipose tissue. Int J Endocrinol 2017:2945012
Wang QA, Tao C, Gupta RK, Scherer PE (2013) Tracking adipogenesis during white adipose tissue development, expansion and regeneration. Nat Med 19:1338–1344
Watts SW, Dorrance AM, Penfold ME, Rourke JL, Sinal CJ, Seitz B, Sullivan TJ, Charvat TT, Thompson JM, Burnett R, Fink GD (2013) Chemerin connects fat to arterial contraction. Arterioscler Thromb Vasc Biol 33:1320–1328
Wei H, Chiba S, Moriwaki C, Kitamura H, Ina K, Aosa T, Tomonari K, Gotoh K, Masaki T, Katsuragi I, Noguchi H, Kakuma T, Hamaguchi K, Shimada T, Fujikura Y, Shibata H (2015) A clinical approach to brown adipose tissue in the para-aortic area of the human thorax. PLoS ONE 10:e0122594
Zhao Y, Waldman SD, Flynn LE (2015) Multilineage co-culture of adipose-derived stem cells for tissue engineering. J Tissue Eng Regen Med 9:826–837
Acknowledgements
This research was supported by NHLBI 5R01HL117847-02 and 2P01HL070687-11A1. The authors acknowledge the technical assistance of Drs. Rahul Nelli and Clarissa Strieder-Barboza at the Department of Large Animal Clinical Sciences and Janice Thompson at the Watts Laboratory, Department of Pharmacology and Toxicology.
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All animal procedures were approved by the Michigan State University Animal Care and Use Committee Procedures.
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Thelen, K., Watts, S.W. & Contreras, G.A. Adipogenic potential of perivascular adipose tissue preadipocytes is improved by coculture with primary adipocytes. Cytotechnology 70, 1435–1445 (2018). https://doi.org/10.1007/s10616-018-0238-0
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DOI: https://doi.org/10.1007/s10616-018-0238-0