Abstract
White adipose tissue (WAT) has attracted interest for tissue engineering and cell-based therapies as an abundant source of adipose stem/stromal cells (ASC). However, technical challenges in WAT cell culture have limited its applications in regenerative medicine. Traditional two-dimensional (2D) cell culture models, which are essentially monolayers of cells on glass or plastic substrates, inadequately represent tissue architecture, biochemical concentration gradients, substrate stiffness, and most importantly for WAT research, cell phenotypic heterogeneity. Physiological cell culture platforms for WAT modeling must recapitulate the native diversity of cell types and their coordination within the organ. For this purpose, we developed a three-dimensional (3D) model using magnetic levitation. Here, we describe our protocol that we successfully employed to build adipose tissue organoids (adipospheres) that preserve the heterogeneity of the constituent cell types in vitro. We demonstrate the capacity of assembling adipospheres from multiple cell types, including ASCs, endohtelial cells, and leukocytes that recreate tissue organization. These adipospheres mimicked WAT organogenesis in that they enabled the formation of vessel-like endothelial structures with lumens and differentiation of unilocular adipocytes. Altogether, magnetic levitation is a cell culture platform that recreates tissue structure, function, and heterogeneity in vitro, and serves as a foundation for high-throughput WAT tissue culture and analysis.
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References
Daquinag AC, Zhang Y, Kolonin MG (2011) Vascular targeting of adipose tissue as an anti-obesity approach. Trends Pharmacol Sci 32(5):300–307
Han J, Koh YJ, Moon HR et al (2010) Adipose tissue is an extramedullary reservoir for functional hematopoietic stem and progenitor cells. Blood 115:957–964
Bianco P (2014) “Mesenchymal” stem cells. Annu Rev Cell Dev Biol 30:677–704
Bianco P, Robey PG, Simmons PJ (2008) Mesenchymal stem cells: revisiting history, concepts, and assays. Cell Stem Cell 2:313–319
Kolonin MG, Simmons PJ (2009) Combinatorial stem cell mobilization. Nat Biotechnol 27(3):252–253
Choi JH, Gimble JM, Lee K et al (2010) Adipose tissue engineering for soft tissue regeneration. Tissue Eng Part B Rev 16:413–426
Scherberich A, Müller AM, Schäfer DJ et al (2010) Adipose tissue-derived progenitors for engineering osteogenic and vasculogenic grafts. J Cell Physiol 225(2):348–353
Güven S, Mehrkens A, Saxer F et al (2011) Engineering of large osteogenic grafts with rapid engraftment capacity using mesenchymal and endothelial progenitors from human adipose tissue. Biomaterials 32:5801–5809
Kolonin MG, Evans KW, Mani SA et al (2012) Alternative origins of stroma in normal organs and disease. Stem Cell Res 8(2):312–323
Schindler M, Nur-E-Kamal A, Ahmed I et al (2006) Living in three dimensions: 3D nanostructured environments for cell culture and regenerative medicine. Cell Biochem Biophys 45:215–227
Griffith LG, Swartz MA (2006) Capturing complex 3D tissue physiology in vitro. Nat Rev Mol Cell Biol 7:211–224
Peyton SR, Kim PD, Ghajar CM et al (2008) The effects of matrix stiffness and RhoA on the phenotypic plasticity of smooth muscle cells in a 3-D biosynthetic hydrogel system. Biomaterials 29:2597–2607
Pedersen JA, Swartz MA (2005) Mechanobiology in the third dimension. Ann Biomed Eng 33:1469–1490
Zhang S (2004) Beyond the Petri dish. Nat Biotechnol 22:151–152
Kleinman HK, Philp D, Hoffman MP (2003) Role of the extracellular matrix in morphogenesis. Curr Opin Biotechnol 14:526–532
Cukierman E, Pankov R, Stevens DR et al (2001) Taking cell-matrix adhesions to the third dimension. Science 294:1708–1712
Pampaloni F, Reynaud EG, Stelzer EHK (2007) The third dimension bridges the gap between cell culture and live tissue. Nat Rev Mol Cell Biol 8:839–845
Zuk PA, Zhu M, Mizuno H et al (2001) Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 7:211–228
Daquinag AC, Zhang Y, Amaya-Manzanares F et al (2011) An isoform of decorin is a resistin receptor on the surface of adipose progenitor cells. Cell Stem Cell 9:74–86
Traktuev DO, Merfeld-Clauss S, Li J et al (2008) A population of multipotent CD34-positive adipose stromal cells share pericyte and mesenchymal surface markers, reside in a periendothelial location, and stabilize endothelial networks. Circ Res 102:77–85
Souza GR, Molina JR, Raphael RM et al (2010) Three-dimensional tissue culture based on magnetic cell levitation. Nat Nanotechnol 5:291–296
Tseng H, Gage JA, Raphael RM et al (2013) Assembly of a three-dimensional multitype bronchiole coculture model using magnetic levitation. Tissue Eng Part C Methods 19:665–675
Molina JR, Hayashi Y, Stephens C et al (2010) Invasive glioblastoma cells acquire stemness and increased Akt activation. Neoplasia 12:453–463
Xu L, Gao G, Ren J et al (2012) Estrogen receptor β of host promotes the progression of lung cancer brain metastasis of an orthotopic mouse model. J Cancer Ther 3:352–358
Lee JS, Morrisett JD, Tung C-H (2012) Detection of hydroxyapatite in calcified cardiovascular tissues. Atherosclerosis 224:340–347
Castro-Chavez F, Vickers KC, Lee JS et al (2013) Effect of lyso-phosphatidylcholine and Schnurri-3 on osteogenic transdifferentiation of vascular smooth muscle cells to calcifying vascular cells in 3D culture. Biochim Biophys Acta 1830:3828–3834
Becker JL, Souza GR (2013) Using space-based investigations to inform cancer research on Earth. Nat Rev Cancer 13:315–327
Haisler WL, Timm DM, Gage JA et al (2013) Three-dimensional cell culturing by magnetic levitation. Nat Protoc 8:1940–1949
Timm DM, Chen J, Sing D et al (2013) A high-throughput three-dimensional cell migration assay for toxicity screening with mobile device-based macroscopic image analysis. Sci Rep 3:3000
Tseng H, Balaoing LR, Grigoryan B et al (2014) A three-dimensional co-culture model of the aortic valve using magnetic levitation. Acta Biomater 10:173–182
Jaganathan H, Gage J, Leonard F et al (2014) Three-dimensional in vitro co-culture model of breast tumor using magnetic levitation. Sci Rep 4:6468
Daquinag AC, Souza GR, Kolonin MG (2013) Adipose tissue engineering in three-dimensional levitation tissue culture system based on magnetic nanoparticles. Tissue Eng Part C Methods 19:336–344
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Tseng, H., Daquinag, A.C., Souza, G.R., Kolonin, M.G. (2018). Three-Dimensional Magnetic Levitation Culture System Simulating White Adipose Tissue. In: Bunnell, B.A., Gimble, J.M. (eds) Adipose-Derived Stem Cells. Methods in Molecular Biology, vol 1773. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7799-4_12
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DOI: https://doi.org/10.1007/978-1-4939-7799-4_12
Publisher Name: Humana Press, New York, NY
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