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Preparation and Characterization of Human Adipose Tissue-Derived Extracellular Matrix, Growth Factors, and Stem Cells: A Concise Review

  • So Young Chun
  • Jeong Ok Lim
  • Eun Hye Lee
  • Man-Hoon Han
  • Yun-Sok Ha
  • Jun Nyung Lee
  • Bum Soo Kim
  • Min Jeong Park
  • MyungGu Yeo
  • Bongsu JungEmail author
  • Tae Gyun KwonEmail author
Review Article
  • 24 Downloads

Abstract

Background:

Human adipose tissue is routinely discarded as medical waste. However, this tissue may have valuable clinical applications since methods have been devised to effectively isolate adipose-derived extracellular matrix (ECM), growth factors (GFs), and stem cells. In this review, we analyze the literature that devised these methods and then suggest an optimal method based on their characterization results.

Methods:

Methods that we analyze in this article include: extraction of adipose tissue, decellularization, confirmation of decellularization, identification of residual active ingredients (ECM, GFs, and cells), removal of immunogens, and comparing structural/physiological/biochemical characteristics of active ingredients.

Results:

Human adipose ECMs are composed of collagen type I–VII, laminin, fibronectin, elastin, and glycosaminoglycan (GAG). GFs immobilized in GAG include basic fibroblast growth factor (bFGF), transforming growth factor beta 1(TGF-b1), insulin like growth factor 1 (IGF-1), vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF), BMP4 (bone morphogenetic protein 4), nerve growth factor (NGF), hepatocyte growth factor (HGF), and epithermal growth factor (EGF). Stem cells in the stromal-vascular fraction display mesenchymal markers, self-renewal gene expression, and multi-differentiation potential.

Conclusion:

Depending on the preparation method, the volume, biological activity, and physical properties of ECM, GFs, and adipose tissue-derived cells can vary. Thus, the optimal preparation method is dependent on the intended application of the adipose tissue-derived products.

Keywords

Human adipose tissue Extracellular matrix Growth factors Adipose-derived stem cell Optimum method 

Notes

Acknowledgements

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) and funded by the Korean government (MSIT) (2014M3A9D3034164), (2016R1C1B1011180), (2018R1C1B5040264) (2019R1A2C1004046), and the Ministry of Trade, Industry and Energy (MOTIE), Korea, under the “Regional Industry Infrastructure and R&D Support Program” (R0005886) supervised by the Korea Institute for Advancement of Technology (KIAT), and 2017–2019 Medical Cluster R&D Support Project through Daegu Gyeongbuk Medical Innovation Foundation funded by the Ministry of Health & Welfare, the Republic of Korea (HI16C2505).

Compliance with ethical standards

Conflict of interest

All authors declare that there is no conflict of interest.

Ethical statement

There are no animal experiments carried out for this article.

References

  1. 1.
    Gomillion CT, Burg KJ. Stem cells and adipose tissue engineering. Biomaterials. 2006;27:6052–63.Google Scholar
  2. 2.
    Patrick CW Jr. Tissue engineering strategies for adipose tissue repair. Anat Rec. 2001;263:361–6.Google Scholar
  3. 3.
    Banyard DA, Borad V, Amezcua E, Wirth GA, Evans GR, Widgerow AD. Preparation, characterization, and clinical implications of human decellularized adipose tissue extracellular matrix (hDAM): a comprehensive review. Aesthet Surg J. 2016;36:349–57.Google Scholar
  4. 4.
    Zhou Q, Desta T, Fenton M, Graves DT, Amar S. Cytokine profiling of macrophages exposed to Porphyromonas gingivalis, its lipopolysaccharide, or its FimA protein. Infect Immun. 2005;73:935–43.Google Scholar
  5. 5.
    Trayhurn P, Wood IS. Signalling role of adipose tissue: adipokines and inflammation in obesity. Biochem Soc Trans. 2005;33:1078–81.Google Scholar
  6. 6.
    Young DA, Christman KL. Injectable biomaterials for adipose tissue engineering. Biomed Mater. 2012;7:024104.Google Scholar
  7. 7.
    Chun SY, Oh SH, Yoo JJ, Kwon TG. Fabrication and characterization techniques for decellularized organ scaffolds. Tissue Eng Regen Med. 2014;11:S1–10.Google Scholar
  8. 8.
    Gentile P, Piccinno MS, Calabrese C. Characteristics and potentiality of human adipose-derived stem cells (hASCs) obtained from enzymatic digestion of fat graft. Cells. 2019;8:E282.Google Scholar
  9. 9.
    Dufrane D. Impact of age on human adipose stem cells for bone tissue engineering. Cell Transplant. 2017;26:1496–504.Google Scholar
  10. 10.
    Brown BN, Freund JM, Han L, Rubin JP, Reing JE, Jeffries EM, et al. Comparison of three methods for the derivation of a biologic scaffold composed of adipose tissue extracellular matrix. Tissue Eng Part C Methods. 2011;17:411–21.Google Scholar
  11. 11.
    Wu I, Nahas Z, Kimmerling KA, Rosson GD, Elisseeff JH. An injectable adipose matrix for soft-tissue reconstruction. Plast Reconstr Surg. 2012;129:1247–57.Google Scholar
  12. 12.
    Choi JS, Yang HJ, Kim BS, Kim JD, Kim JY, Yoo B, et al. Human extracellular matrix (ECM) powders for injectable cell delivery and adipose tissue engineering. J Control Release. 2009;139:2–7.Google Scholar
  13. 13.
    Choi YC, Choi JS, Kim BS, Kim JD, Yoon HI, Cho YW. Decellularized extracellular matrix derived from porcine adipose tissue as a xenogeneic biomaterial for tissue engineering. Tissue Eng Part C Methods. 2012;18:866–76.Google Scholar
  14. 14.
    Shi Y, Dittman B, Atkinson B, Stilwell RL. Cell repopulated collagen matrix for soft tissue repair and regeneration. US Patent 20140286911A1, 9 Sept 2014 (2014).Google Scholar
  15. 15.
    Song M, Liu Y, Hui L. Preparation and characterization of acellular adipose tissue matrix using a combination of physical and chemical treatments. Mol Med Rep. 2018;17:138–46.Google Scholar
  16. 16.
    Choi JS, Kim BS, Kim JY, Kim JD, Choi YC, Yang HJ, et al. Decellularized extracellular matrix derived from human adipose tissue as a potential scaffold for allograft tissue engineering. J Biomed Mater Res A. 2011;97:292–9.Google Scholar
  17. 17.
    Gilbert TW, Sellaro TL, Badylak SF. Decellularization of tissues and organs. Biomaterials. 2006;27:3675–83.Google Scholar
  18. 18.
    Gilbert TW, Freund JM, Badylak SF. Quantification of DNA in biologic scaffold materials. J Surg Res. 2009;152:135–9.Google Scholar
  19. 19.
    Flynn LE. The use of decellularized adipose tissue to provide an inductive microenvironment for the adipogenic differentiation of human adipose-derived stem cells. Biomaterials. 2010;31:4715–24.Google Scholar
  20. 20.
    Rieder E, Kasimir MT, Silberhumer G, Seebacher G, Wolner E, Simon P, et al. Decellularization protocols of porcine heart valves differ importantly in efficiency of cell removal and susceptibility of the matrix to recellularization with human vascular cells. J Thorac Cardiovasc Surg. 2004;127:399–405.Google Scholar
  21. 21.
    Chung YC, Su YP, Chen CC, Jia G, Wang HL, Wu JC, et al. Relationship between antibacterial activity of chitosan and surface characteristics of cell wall. Acta Pharmacol Sin. 2004;25:932–6.Google Scholar
  22. 22.
    Choi JS, Yang HJ, Kim BS, Kim JD, Lee SH, Lee EK, et al. Fabrication of porous extracellular matrix scaffolds from human adipose tissue. Tissue Eng Part C Methods. 2010;16:387–96.Google Scholar
  23. 23.
    Ricard-Blum S. The collagen family. Cold Spring Harb Perspect Biol. 2011;3:a004978.Google Scholar
  24. 24.
    Chun TH. Peri-adipocyte ECM remodeling in obesity and adipose tissue fibrosis. Adipocyte. 2012;1:89–95.Google Scholar
  25. 25.
    Streuli C. Extracellular matrix remodelling and cellular differentiation. Curr Opin Cell Biol. 1999;11:634–40.Google Scholar
  26. 26.
    Niyibizi C, Fietzek PP, van der Rest M. Human placenta type V collagens. Evidence for the existence of an alpha 1(V) alpha 2(V) alpha 3(V) collagen molecule. J Biol Chem. 1984;259:14170–4.Google Scholar
  27. 27.
    Noro A, Sillat T, Virtanen I, Ingerpuu S, Bäck N, Konttinen YT, et al. Laminin production and basement membrane deposition by mesenchymal stem cells upon adipogenic differentiation. J Histochem Cytochem. 2013;61:719–30.Google Scholar
  28. 28.
    Atkinson JJ, Adair-Kirk TL, Kelley DG, Demello D, Senior RM. Clara cell adhesion and migration to extracellular matrix. Respir Res. 2008;9:1.Google Scholar
  29. 29.
    Spiegelman BM, Ginty CA. Fibronectin modulation of cell shape and lipogenic gene expression in 3T3-adipocytes. Cell. 1983;35:657–66.Google Scholar
  30. 30.
    Mosher DF, Schad PE. Cross-linking of fibronectin to collagen by blood coagulation Factor XIIIa. J Clin Invest. 1979;64:781–7.Google Scholar
  31. 31.
    Khan T, Muise ES, Iyengar P, Wang ZV, Chandalia M, Abate N, et al. Metabolic dysregulation and adipose tissue fibrosis: role of collagen VI. Mol Cell Biol. 2009;29:1575–91.Google Scholar
  32. 32.
    Spencer M, Unal R, Zhu B, Rasouli N, McGehee RE Jr, Peterson CA, et al. Adipose tissue extracellular matrix and vascular abnormalities in obesity and insulin resistance. J Clin Endocrinol Metab. 2011;96:E1990–8.Google Scholar
  33. 33.
    Martinez-Santibanez G, Singer K, Cho KW, DelProposto JL, Mergian T, Lumeng CN. Obesity-induced remodeling of the adipose tissue elastin network is independent of the metalloelastase MMP-12. Adipocyte. 2015;4:264–72.Google Scholar
  34. 34.
    Kielty CM, Sherratt MJ, Shuttleworth CA. Elastic fibres. J Cell Sci. 2002;115:2817–28.Google Scholar
  35. 35.
    Holst J, Watson S, Lord MS, Eamegdool SS, Bax DV, Nivison-Smith LB, et al. Substrate elasticity provides mechanical signals for the expansion of hemopoietic stem and progenitor cells. Nat Biotechnol. 2010;28:1123–8.Google Scholar
  36. 36.
    Doran MR, Markway BD, Aird IA, Rowlands AS, George PA, Nielsen LK, et al. Surface-bound stem cell factor and the promotion of hematopoietic cell expansion. Biomaterials. 2009;30:4047–52.Google Scholar
  37. 37.
    Mullen LM, Best SM, Brooks RA, Ghose S, Gwynne JH, Wardale J, et al. Binding and release characteristics of insulin-like growth factor-1 from a collagen-glycosaminoglycan scaffold. Tissue Eng Part C Methods. 2010;16:1439–48.Google Scholar
  38. 38.
    Choi JS, Kim BS, Kim JD, Choi YC, Lee HY, Cho YW. In vitro cartilage tissue engineering using adipose-derived extracellular matrix scaffolds seeded with adipose-derived stem cells. Tissue Eng Part A. 2012;18:80–92.Google Scholar
  39. 39.
    Raeder RH, Badylak SF, Sheehan C, Kallakury B, Metzger DW. Natural anti-galactose alpha1,3 galactose antibodies delay, but do not prevent the acceptance of extracellular matrix xenografts. Transpl Immunol. 2002;10:15–24.Google Scholar
  40. 40.
    Daly KA, Stewart-Akers AM, Hara H, Ezzelarab M, Long C, Cordero K, et al. Effect of the alphaGal epitope on the response to small intestinal submucosa extracellular matrix in a nonhuman primate model. Tissue Eng Part A. 2009;15:3877–88.Google Scholar
  41. 41.
    van Dongen JA, Getova V, Brouwer LA, Liguori GR, Sharma PK, Stevens HP, et al. Adipose tissue-derived extracellular matrix hydrogels as a release platform for secreted paracrine factors. J Tissue Eng Regen Med. 2019;13:973–85.Google Scholar
  42. 42.
    Lin P, Chan WC, Badylak SF, Bhatia SN. Assessing porcine liver-derived biomatrix for hepatic tissue engineering. Tissue Eng. 2004;10:1046–53.Google Scholar
  43. 43.
    Badylak SF, Freytes DO, Gilbert TW. Extracellular matrix as a biological scaffold material: structure and function. Acta Biomater. 2009;5:1–13.Google Scholar
  44. 44.
    Kokai LE, Schilling BK, Chnari E, Huang YC, Imming EA, Karunamurthy A, et al. Injectable allograft adipose matrix supports adipogenic tissue remodeling in the nude mouse and human. Plast Reconstr Surg. 2019;143:299e–309.Google Scholar
  45. 45.
    Young DA, Ibrahim DO, Hu D, Christman KL. Injectable hydrogel scaffold from decellularized human lipoaspirate. Acta Biomater. 2011;7:1040–9.Google Scholar
  46. 46.
    Shan X, Choi JH, Kim KJ, Lee YJ, Ryu YH, Lee SJ, et al. Adipose stem cells with conditioned media for treatment of acne vulgaris scar. Tissue Eng Regen Med. 2018;15:49–61.Google Scholar
  47. 47.
    Saksela O, Moscatelli D, Sommer A, Rifkin DB. Endothelial cell-derived heparan sulfate binds basic fibroblast growth factor and protects it from proteolytic degradation. J Cell Biol. 1988;107:743–51.Google Scholar
  48. 48.
    Santana H, González Y, Campana PT, Noda J, Amarantes O, Itri R, et al. Screening for stability and compatibility conditions of recombinant human epidermal growth factor for parenteral formulation: effect of pH, buffers, and excipients. Int J Pharm. 2013;452:52–62.Google Scholar
  49. 49.
    Senderoff RI, Wootton SC, Boctor AM, Chen TM, Giordani AB, Julian TN, et al. Aqueous stability of human epidermal growth factor 1-48. Pharm Res. 1994;11:1712–20.Google Scholar
  50. 50.
    Elias AP, Dias S. Microenvironment changes (in pH) affect VEGF alternative splicing. Cancer Microenviron. 2008;1:131–9.Google Scholar
  51. 51.
    Hoener MC, Varon S. Effects of sodium chloride, Triton X-100, and alkaline pH on the measurable contents and sedimentability of the nerve growth factor (NGF) antigen in adult rat hippocampal tissue extracts. J Neurosci Res. 1997;49:508–14.Google Scholar
  52. 52.
    Dong X, Wei X, Yi W, Gu C, Kang X, Liu Y, et al. RGD-modified acellular bovine pericardium as a bioprosthetic scaffold for tissue engineering. J Mater Sci Mater Med. 2009;20:2327–36.Google Scholar
  53. 53.
    Wong ML, Wong JL, Horn RM, Sannajust KC, Rice DA, Griffiths LG. Effect of urea and thiourea on generation of xenogeneic extracellular matrix scaffolds for tissue engineering. Tissue Eng part C Methods. 2016;22:700–7.Google Scholar
  54. 54.
    Vavken P, Joshi S, Murray MM. TRITON-X is most effective among three decellularization agents for ACL tissue engineering. J Orthop Res. 2009;27:1612–8.Google Scholar
  55. 55.
    Niemelä S, Miettinen S, Sarkanen JR, Ashammakhi N. Adipose tissue and adipocyte differentiation: molecular and cellular aspects and tissue engineering applications. In: Ashammakhi N, Reis R, Chiellini F, editors. Topics in tissue engineering. Vol. 4; 2008. Chap. 4. p. 1–26.Google Scholar
  56. 56.
    Florini JR, Magri KA. Effects of growth factors on myogenic differentiation. Am J Physiol. 1989;256:C701–11.Google Scholar
  57. 57.
    Giustina A, Mazziotti G, Canalis E. Growth hormone, insulin-like growth factors, and the skeleton. Endocr Rev. 2008;29:535–59.Google Scholar
  58. 58.
    Aberg D. Role of the growth hormone/insulin-like growth factor 1 axis in neurogenesis. Endocr Dev. 2010;17:63–76.Google Scholar
  59. 59.
    von Heimburg D, Serov G, Oepen T, Pallua N. Fat tissue engineering. In: Ashammakhi N, Ferretti P, editors. Topics in tissue engineering. 2003. Chap. 8. p. 1–16.Google Scholar
  60. 60.
    Park JH, Kim KJ, Rhie JW, Oh IH. Characterization of adipose tissue mesenchymal stromal cell subsets with distinct plastic adherence. Tissue Eng Regen Med. 2016;13:39–46.Google Scholar
  61. 61.
    Bianco P, Robey PG, Simmons PJ. Mesenchymal stem cells: revisiting history, concepts, and assays. Cell Stem Cell. 2008;2:313–9.Google Scholar
  62. 62.
    Maurer MH. Proteomic definitions of mesenchymal stem cells. Stem Cells Int. 2011;2011:704256.Google Scholar
  63. 63.
    Schmelzer E, McKeel DT, Gerlach JC. Characterization of human mesenchymal stem cells from different tissues and their membrane encasement for prospective transplantation therapies. Biomed Res Int. 2019;2019:6376271.Google Scholar
  64. 64.
    Salgado AJ, Reis RL, Sousa NJ, Gimble JM. Adipose tissue derived stem cells secretome: soluble factors and their roles in regenerative medicine. Curr Stem Cell Res Ther. 2010;5:103–10.Google Scholar
  65. 65.
    Dmitrieva LA, Elizova LA. Structural changes in human dentin with the use of modern filling materials. Stomatologiia (Mosk). 1991;5:21–3.Google Scholar
  66. 66.
    Strem BM, Zhu M, Alfonso Z, Daniels EJ, Schreiber R, Beygui R, et al. Expression of cardiomyocytic markers on adipose tissue-derived cells in a murine model of acute myocardial injury. Cytotherapy. 2005;7:282–91.Google Scholar
  67. 67.
    Schäffler A, Büchler C. Concise review: adipose tissue-derived stromal cells–basic and clinical implications for novel cell-based therapies. Stem Cells. 2007;25:818–27.Google Scholar
  68. 68.
    Shenaq DS, Rastegar F, Petkovic D, Zhang BQ, He BC, Chen L, et al. Mesenchymal progenitor cells and their orthopedic applications: forging a path towards clinical trials. Stem Cells Int. 2010;2010:519028.Google Scholar
  69. 69.
    Murata D, Akieda S, Misumi K, Nakayama K. Osteochondral regeneration with a scaffold-free three-dimensional construct of adipose tissue-derived mesenchymal stromal cells in pigs. Tissue Eng Regen Med. 2018;15:101–13.Google Scholar
  70. 70.
    Dicker A, Le Blanc K, Aström G, van Harmelen V, Götherström C, Blomqvist L, et al. Functional studies of mesenchymal stem cells derived from adult human adipose tissue. Exp Cell Res. 2005;308:283–90.Google Scholar
  71. 71.
    Mori S, Kiuchi S, Ouchi A, Hase T, Murase T. Characteristic expression of extracellular matrix in subcutaneous adipose tissue development and adipogenesis; comparison with visceral adipose tissue. Int J Biol Sci. 2014;10:825–33.Google Scholar
  72. 72.
    Virtue S, Vidal-Puig A. Adipose tissue expandability, lipotoxicity and the metabolic syndrome: an allostatic perspective. Biochim Biophys Acta. 2010;1801:338–49.Google Scholar
  73. 73.
    Mariman EC, Wang P. Adipocyte extracellular matrix composition, dynamics and role in obesity. Cell Mol Life Sci. 2010;67:1277–92.Google Scholar
  74. 74.
    Herrero L, Shapiro H, Nayer A, Lee J, Shoelson SE. Inflammation and adipose tissue macrophages in lipodystrophic mice. Proc Natl Acad Sci U S A. 2010;107:240–5.Google Scholar
  75. 75.
    Zheng W, McLerran DF, Rolland B, Zhang X, Inoue M, Matsuo K, et al. Association between body-mass index and risk of death in more than 1 million Asians. N Engl J Med. 2011;364:719–29.Google Scholar
  76. 76.
    Berrington de Gonzalez A, Hartge P, Cerhan JR, Flint AJ, Hannan L, MacInnis RJ, et al. Body-mass index and mortality among 1.46 million white adults. N Engl J Med. 2010;363:2211–9.Google Scholar
  77. 77.
    Ministry of Food and drug safety of KOREA. Guidelines for evaluating cell therapeutic agents including scaffolds. B1-2014-3-011. 2014.Google Scholar
  78. 78.
    Noverina R, Widowati W, Ayuningtyas W, Kurniawan D, Afifah E, Laksmitawati DR, et al. Growth factors profile in conditioned medium human adipose tissue-derived mesenchymal stem cells (CM-hATMSCs). Clin Nutr Exp. 2019;24:34–44.Google Scholar

Copyright information

© The Korean Tissue Engineering and Regenerative Medicine Society 2019

Authors and Affiliations

  1. 1.BioMedical Research Institute, Joint Institute for Regenerative MedicineKyungpook National University HospitalDaeguRepublic of Korea
  2. 2.Department of Pathology, School of MedicineKyungpook National UniversityDaeguRepublic of Korea
  3. 3.Department of Urology, School of MedicineKyungpook National UniversityDaeguRepublic of Korea
  4. 4.Medical Device Development CenterDaegu-Gyeongbuk Medical Innovation Foundation (DGMIF)DaeguRepublic of Korea
  5. 5.Department of UrologyKyungpook National University Chilgok HospitalDaeguRepublic of Korea

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