Analysis of in vitro secretion profiles from adipose-derived cell populations

Journal of Translational Medicine

Benjamin R Herbert¹  Graham Vesey³  Donald Kuah⁴  Edmond J Breen²  Cameron J Hill¹  Rebecca A Webster³  Sinead P Blaber³ 

[1] Department of Chemistry and Biomolecular Sciences, Macquarie University, Office 256, Building E8C, Balaclava Rd, North Ryde, NSW 2109, Australia;Australian Proteome Analysis Facility, Macquarie University, North Ryde, NSW, Australia;Regeneus Ltd, Gordon, NSW, Australia;Sydney Sportmed Specialists, Sydney, NSW, Australia

关键词: Bio-Plex;    Growth factors;    Cytokines;    Stromal vascular fraction;    Adipose-derived stem cells;    Adipose tissue;   

Others  :  1205909
DOI  :  10.1186/1479-5876-10-172

 received in 2012-07-01, accepted in 2012-08-16,  发布年份 2012

【 摘 要 】

Background

Adipose tissue is an attractive source of cells for therapeutic purposes because of the ease of harvest and the high frequency of mesenchymal stem cells (MSCs). Whilst it is clear that MSCs have significant therapeutic potential via their ability to secrete immuno-modulatory and trophic cytokines, the therapeutic use of mixed cell populations from the adipose stromal vascular fraction (SVF) is becoming increasingly common.

Methods

In this study we have measured a panel of 27 cytokines and growth factors secreted by various combinations of human adipose-derived cell populations. These were 1. co-culture of freshly isolated SVF with adipocytes, 2. freshly isolated SVF cultured alone, 3. freshly isolated adipocytes alone and 4. adherent adipose-derived mesenchymal stem cells (ADSCs) at passage 2. In addition, we produced an ‘in silico’ dataset by combining the individual secretion profiles obtained from culturing the SVF with that of the adipocytes. This was compared to the secretion profile of co-cultured SVF and adipocytes. Two-tailed t-tests were performed on the secretion profiles obtained from the SVF, adipocytes, ADSCs and the ‘in silico’ dataset and compared to the secretion profiles obtained from the co-culture of the SVF with adipocytes. A p-value of < 0.05 was considered statistically different. To assess the overall changes that may occur as a result of co-culture we compared the proteomes of SVF and SVF co-cultured with adipocytes using iTRAQ quantitative mass spectrometry.

Results

A co-culture of SVF and adipocytes results in a distinct secretion profile when compared to all other adipose-derived cell populations studied. This illustrates that cellular crosstalk during co-culture of the SVF with adipocytes modulates the production of cytokines by one or more cell types. No biologically relevant differences were detected in the proteomes of SVF cultured alone or co-cultured with adipocytes.

Conclusions

The use of mixed adipose cell populations does not appear to induce cellular stress and results in enhanced secretion profiles. Given the importance of secreted cytokines in cell therapy, the use of a mixed cell population such as the SVF with adipocytes may be considered as an alternative to MSCs or fresh SVF alone.

【 授权许可】

   
2012 Blaber et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1 .

Figure 2 .

Figure 3 .

Figure 4 .

Figure 5 .

Figure 6 .

【 参考文献 】

• [1]Trayhurn P, Wood IS: Adipokines: inflammation and the pleiotropic role of white adipose tissue. Br J Nutr 2004, 92:347-355.
• [2]Varma MJ, Breuls RG, Schouten TE, Jurgens WJ, Bontkes HJ, Schuurhuis GJ, van Ham SM, van Milligen FJ: Phenotypical and functional characterization of freshly isolated adipose tissue-derived stem cells. Stem Cell Dev 2007, 16:91-104.
• [3]Riordan NH, Ichim TE, Min WP, Wang H, Solano F, Lara F, Alfaro M, Rodriguez JP, Harman RJ, Patel AN, et al.: Non-expanded adipose stromal vascular fraction cell therapy for multiple sclerosis. J Translational Med 2009., 7
• [4]Astori G, Vignati F, Bardelli S, Tubio M, Gola M, Albertini V, Bambi F, Scali G, Castelli D, Rasini V, et al.: "In vitro" and multicolor phenotypic characterization of cell subpopulations identified in fresh human adipose tissue stromal vascular fraction and in the derived mesenchymal stem cells. J Translational Med 2007, 5:55. BioMed Central Full Text
• [5]Fraser JK, Wulur I, Alfonso Z, Hedrick MH: Fat tissue: an underappreciated source of stem cells for biotechnology. Trends Biotechnol 2006, 24:150-154.
• [6]Strem BM, Hicok KC, Zhu M, Wulur I, Alfonso Z, Schreiber RE, Fraser JK, Hedrick MH: Multipotential differentiation of adipose tissue-derived stem cells. Keio J Med 2005, 54:132-141.
• [7]Estes BT, Wu AW, Guilak F: Potent induction of chondrocytic differentiation of human adipose-derived adult stem cells by bone morphogenetic protein 6. Arthritis Rheum 2006, 54:1222-1232.
• [8]da Silva Meirelles L, Sand TT, Harman RJ, Lennon DP, Caplan AI: MSC frequency correlates with blood vessel density in equine adipose tissue. Tissue Eng Part A 2009, 15:221-229.
• [9]Corcione A, Benvenuto F, Ferretti E, Giunti D, Cappiello V, Cazzanti F, Risso M, Gualandi F, Mancardi GL, Pistoia V, Uccelli A: Human mesenchymal stem cells modulate B-cell functions. Blood 2006, 107:367-372.
• [10]Di Nicola M, Carlo-Stella C, Magni M, Milanesi M, Longoni PD, Matteucci P, Grisanti S, Gianni AM: Human bone marrow stromal cells suppress T-lymphocyte proliferation induced by cellular or nonspecific mitogenic stimuli. Blood 2002, 99:3838-3843.
• [11]Krampera M, Glennie S, Dyson J, Scott D, Laylor R, Simpson E, Dazzi F: Bone marrow mesenchymal stem cells inhibit the response of naive and memory antigen-specific T cells to their cognate peptide. Blood 2003, 101:3722-3729.
• [12]Tse WT, Pendleton JD, Beyer WM, Egalka MC, Guinan EC: Suppression of allogeneic T-cell proliferation by human marrow stromal cells: implications in transplantation. Transplant 2003, 75:389-397.
• [13]Bartholomew A, Sturgeon C, Siatskas M, Ferrer K, McIntosh K, Patil S, Hardy W, Devine S, Ucker D, Deans R, et al.: Mesenchymal stem cells suppress lymphocyte proliferation in vitro and prolong skin graft survival in vivo. Exp Hematol 2002, 30:42-48.
• [14]Aggarwal S, Pittenger MF: Human mesenchymal stem cells modulate allogeneic immune cell responses. Blood 2005, 105:1815-1822.
• [15]Jiang XX, Zhang Y, Liu B, Zhang SX, Wu Y, Yu XD, Mao N: Human mesenchymal stem cells inhibit differentiation and function of monocyte-derived dendritic cells. Blood 2005, 105:4120-4126.
• [16]Meirelles Lda S, Fontes AM, Covas DT, Caplan AI: Mechanisms involved in the therapeutic properties of mesenchymal stem cells. Cytokine Growth Factor Rev 2009, 20:419-427.
• [17]Mahalatchimy A, Rial-Sebbag E, Tournay V, Faulkner A: The Legal Landscape for Advanced Therapies: Material and Institutional Implementation of European Union Rules in France and the United Kingdom. J Law Soc 2012, 39:131-149.
• [18]Nishimura S, Manabe I, Nagasaki M, Eto K, Yamashita H, Ohsugi M, Otsu M, Hara K, Ueki K, Sugiura S, et al.: CD8+ effector T cells contribute to macrophage recruitment and adipose tissue inflammation in obesity. Nat Med 2009, 15:914-920.
• [19]Poggi M, Jager J, Paulmyer-Lacroix O, Peiretti F, Gremeaux T, Verdier M, Grino M, Stepanian A, Msika S, Burcelin R, et al.: The inflammatory receptor CD40 is expressed on human adipocytes: contribution to crosstalk between lymphocytes and adipocytes. Diabetologia 2009, 52:1152-1163.
• [20]Suganami T, Nishida J, Ogawa Y: A paracrine loop between adipocytes and macrophages aggravates inflammatory changes - Role of free fatty acids and tumor necrosis factor alpha. Arterioscler Thromb Vasc Biol 2005, 25:2062-2068.
• [21]Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, Benhaim P, Lorenz HP, Hedrick MH: Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 2001, 7:211-228.
• [22]Gorzelniak K, Engeli S, Sharma AM: Standardizing the counting of adipocytes in cell culture. Biotechniques 1998, 24:536-538.
• [23]Fernyhough ME, Vierck JL, Hausman GJ, Mir PS, Okine EK, Dodson MV: Primary adipocyte culture: adipocyte purification methods may lead to a new understanding of adipose tissue growth and development. Cytotechnology 2004, 46:163-172.
• [24]Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, Prockop D, Horwitz E: Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 2006, 8:315-317.
• [25]Biancone L, Bruno S, Deregibus MC, Tetta C, Camussi G: Therapeutic potential of mesenchymal stem cell-derived microvesicles. Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association 2012, 27:3037-3042.
• [26]Permana PA, Menge C, Reaven PD: Macrophage-secreted factors induce adipocyte inflammation and insulin resistance. Biochem Biophys Res Commun 2006, 341:507-514.
• [27]Mitchell JB, McIntosh K, Zvonic S, Garrett S, Floyd ZE, Kloster A, Di Halvorsen Y, Storms RW, Goh B, Kilroy G, et al.: Immunophenotype of human adipose-derived cells: temporal changes in stromal-associated and stem cell-associated markers. Stem Cells 2006, 24:376-385.
• [28]Bunnell BA, Flaat M, Gagliardi C, Patel B, Ripoll C: Adipose-derived stem cells: isolation, expansion and differentiation. Methods 2008, 45:115-120.
• [29]Tansey JT, Sztalryd C, Hlavin EM, Kimmel AR, Londos C: The central role of perilipin A in lipid metabolism and adipocyte lipolysis. IUBMB Life 2004, 56:379-385.
• [30]Davoli R, Gandolfi G, Braglia S, Comella M, Zambonelli P, Buttazzoni L, Russo V: New SNP of the porcine perilipin 2 (PLIN2) gene, association with carcass traits and expression analysis in skeletal muscle. Mol Biol Rep 2011, 38:1575-1583.
• [31]Jernas M, Olsson B, Arner P, Jacobson P, Sjostrom L, Walley A, Froguel P, McTernan PG, Hoffstedt J, Carlsson LM: Regulation of carboxylesterase 1 (CES1) in human adipose tissue. Biochem Biophys Res Commun 2009, 383:63-67.
• [32]Morris NJ, Ducret A, Aebersold R, Ross SA, Keller SR, Lienhard GE: Membrane amine oxidase cloning and identification as a major protein in the adipocyte plasma membrane. J Biol Chem 1997, 272:9388-9392.
• [33]Swierczynski J, Zabrocka L, Goyke E, Raczynska S, Adamonis W, Sledzinski Z: Enhanced glycerol 3-phosphate dehydrogenase activity in adipose tissue of obese humans. Mol Cell Biochem 2003, 254:55-59.
• [34]Berndt J, Kovacs P, Ruschke K, Kloting N, Fasshauer M, Schon MR, Korner A, Stumvoll M, Bluher M: Fatty acid synthase gene expression in human adipose tissue: association with obesity and type 2 diabetes. Diabetologia 2007, 50:1472-1480.
• [35]Janke J, Engeli S, Gorzelniak K, Luft FC, Sharma AM: Mature adipocytes inhibit in vitro differentiation of human preadipocytes via angiotensin type 1 receptors. Diabetes 2002, 51:1699-1707.
• [36]Rosen ED, MacDougald OA: Adipocyte differentiation from the inside out. Nat Rev Mol Cell Biol 2006, 7:885-896.
• [37]Mauney JR, Nguyen T, Gillen K, Kirker-Head C, Gimble JM, Kaplan DL: Engineering adipose-like tissue in vitro and in vivo utilizing human bone marrow and adipose-derived mesenchymal stem cells with silk fibroin 3D scaffolds. Biomaterials 2007, 28:5280-5290.
• [38]Leonard JP, Sherman ML, Fisher GL, Buchanan LJ, Larsen G, Atkins MB, Sosman JA, Dutcher JP, Vogelzang NJ, Ryan JL: Effects of single-dose interleukin-12 exposure on interleukin-12-associated toxicity and interferon-gamma production. Blood 1997, 90:2541-2548.
• [39]Vial T, Descotes J: Immune-mediated side-effects of cytokines in humans. Toxicology 1995, 105:31-57.
• [40]Tayal V, Kalra BS: Cytokines and anti-cytokines as therapeutics–an update. Eur J Pharmacol 2008, 579:1-12.
• [41]Asahara T, Bauters C, Zheng LP, Takeshita S, Bunting S, Ferrara N, Symes JF, Isner JM: Synergistic effect of vascular endothelial growth factor and basic fibroblast growth factor on angiogenesis in vivo. Circulation 1995, 92:II365-II371.
• [42]Carmeliet P, Moons L, Luttun A, Vincenti V, Compernolle V, De Mol M, Wu Y, Bon F, Devy L, Beck H, et al.: Synergism between vascular endothelial growth factor and placental growth factor contributes to angiogenesis and plasma extravasation in pathological conditions. Nat Med 2001, 7:575-583.
• [43]Cao R, Brakenhielm E, Pawliuk R, Wariaro D, Post MJ, Wahlberg E, Leboulch P, Cao Y: Angiogenic synergism, vascular stability and improvement of hind-limb ischemia by a combination of PDGF-BB and FGF-2. Nat Med 2003, 9:604-613.
• [44]Chae JK, Kim I, Lim ST, Chung MJ, Kim WH, Kim HG, Ko JK, Koh GY: Coadministration of angiopoietin-1 and vascular endothelial growth factor enhances collateral vascularization. Arterioscler Thromb Vasc Biol 2000, 20:2573-2578.
• [45]Togel F, Weiss K, Yang Y, Hu ZM, Zhang P, Westenfelder C: Vasculotropic, paracrine actions of infused mesenchymal stem cells are important to the recovery from acute kidney injury. Am J Physiol Renal Physiol 2007, 292:F1626-F1635.
• [46]Caplan AI, Dennis JE: Mesenchymal stem cells as trophic mediators. J Cell Biochem 2006, 98:1076-1084.
• [47]Ortiz LA, Gambelli F, McBride C, Gaupp D, Baddoo M, Kaminski N, Phinney DG: Mesenchymal stem cell engraftment in lung is enhanced in response to bleomycin exposure and ameliorates its fibrotic effects. Proc Natl Acad Sci USA 2003, 100:8407-8411.
• [48]Laskin DL, Sunil VR, Gardner CR, Laskin JD: Macrophages and tissue injury: agents of defense or destruction? Annu Rev Pharmacol Toxicol 2011, 51:267-288.
• [49]Chopp M, Li Y: Treatment of neural injury with marrow stromal cells. Lancet Neurol 2002, 1:92-100.