Enzymatic and non-enzymatic isolation systems for adipose tissue-derived cells: current state of the art

Cell Regeneration - Tập 4 - Trang 1-14 - 2015
Eleni Oberbauer1,2, Carolin Steffenhagen1,2, Christoph Wurzer1,2, Christian Gabriel1,2,3, Heinz Redl1,2, Susanne Wolbank1,2
1Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research Center, Linz/Vienna, Austria
2Austrian Cluster for Tissue Regeneration, Vienna, Austria
3Red Cross Blood Transfusion Service of Upper Austria, Linz, Austria

Tóm tắt

In the past decade, adipose tissue became a highly interesting source of adult stem cells for plastic surgery and regenerative medicine. The isolated stromal vascular fraction (SVF) is a heterogeneous cell population including the adipose-derived stromal/stem cells (ASC), which showed regenerative potential in several clinical studies and trials. SVF should be provided in a safe and reproducible manner in accordance with current good manufacturing practices (cGMP). To ensure highest possible safety for patients, a precisely defined procedure with a high-quality control is required. Hence, an increasing number of adipose tissue-derived cell isolation systems have been developed. These systems aim for a closed, sterile, and safe isolation process limiting donor variations, risk for contaminations, and unpredictability of the cell material. To isolate SVF from adipose tissue, enzymes such as collagenase are used. Alternatively, in order to avoid enzymes, isolation systems using physical forces are available. Here, we provide an overview of known existing enzymatic and non-enzymatic adipose tissue-derived cell isolation systems, which are patented, published, or already on the market.

Tài liệu tham khảo

Coleman 3rd WP, Glogau RG, Klein JA, Moy RL, Narins RS, Chuang TY, et al. Guidelines of care for liposuction. J Am Acad Dermatol. 2001;45(3):438–47. doi:10.1067/mjd.2001.117045. Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, et al. Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng. 2001;7(2):211–28. doi:10.1089/107632701300062859. Harwood Jr HJ. The adipocyte as an endocrine organ in the regulation of metabolic homeostasis. Neuropharmacology. 2012;63(1):57–75. doi:10.1016/j.neuropharm.2011.12.010. Coelho M, Oliveira T, Fernandes R. Biochemistry of adipose tissue: an endocrine organ. Arch Med Sci: AMS. 2013;9(2):191–200. doi:10.5114/aoms.2013.33181. Minguell JJ, Erices A, Conget P. Mesenchymal stem cells. Exp Biol Med (Maywood, NJ). 2001;226(6):507–20. Anker PS I ’t, Scherjon SA, Kleijburg-van der Keur C, de Groot-Swings GM, Claas FH, Fibbe WE, et al. Isolation of mesenchymal stem cells of fetal or maternal origin from human placenta. Stem Cells. 2004;22(7):1338–45. doi:10.1634/stemcells.2004-0058. Fukuchi Y, Nakajima H, Sugiyama D, Hirose I, Kitamura T, Tsuji K. Human placenta-derived cells have mesenchymal stem/progenitor cell potential. Stem Cells. 2004;22(5):649–58. doi:10.1634/stemcells.22-5-649. De Bari C, Dell’Accio F, Tylzanowski P, Luyten F. Multipotent mesenchymal stem cells from adult human synovial membrane. Arthritis Rheum. 2001;44:1928–42. doi:10.1002/1529-0131(200108)44. Young H, Steele T, Bray R, Hudson J, Floyd J, Hawkins K, et al. Human reserve pluripotent mesenchymal stem cells are present in the connective tissues of skeletal muscle and dermis derived from fetal, adult, and geriatric donors. Anat Rec. 2001;264:51–62. Zvaifler N, Marinova-Mutafchieva L, Adams G, Edwards C, Moss J, Burger J, et al. Mesenchymal precursor cells in the blood of normal individuals. Arthritis Res. 2000;2:477–88. Erices A, Conget P, Minguell JJ. Mesenchymal progenitor cells in human umbilical cord blood. Br J Haematol. 2000;109(1):235–42. doi:10.1046/j.1365-2141.2000.01986.x. Wakitani S, Goto T, Pineda S, Young R, Mansour J, Caplan A, et al. Mesenchymal cell-based repair of large, full-thickness defects of articular cartilage. J Bone Joint Surg Am Vol. 1994;76:579–92. Arai F, Ohneda O, Miyamoto T, Zhang XQ, Suda T. Mesenchymal stem cells in perichondrium express activated leukocyte cell adhesion molecule and participate in bone marrow formation. J Exp Med. 2002;195(12):1549–63. doi:10.1084/jem.20011700. Gronthos S, Franklin D, Leddy H, Robey P, Storms R, Gimble J. Surface protein characterization of human adipose tissue-derived stromal cells. J Cell Physiol. 2001;189:54–63. doi:10.1002/jcp.1138. Bailo M, Soncini M, Vertua E, Signoroni PB, Sanzone S, Lombardi G, et al. Engraftment potential of human amnion and chorion cells derived from term placenta. Transplantation. 2004;78(10):1439–48. doi:10.1093/humrep/den356. Pittenger M, Mackay A, Beck S, Jaiswal R, Douglas R, Mosca J, et al. Multilineage potential of adult human mesenchymal stem cells. Science (New York, NY). 1999;284:143–7. doi:10.1126/science.284.5411.143. Tse WT, Pendleton JD, Beyer WM, Egalka MC, Guinan EC. Suppression of allogeneic T-cell proliferation by human marrow stromal cells: implications in transplantation. Transplantation. 2003;75(3):389–97. doi:10.1097/01.tp.0000045055.63901.a9. Puissant B, Barreau C, Bourin P, Clavel C, Corre J, Bousquet C, et al. Immunomodulatory effect of human adipose tissue-derived adult stem cells: comparison with bone marrow mesenchymal stem cells. Br J Haematol. 2005;129(1):118–29. doi:10.1111/j.1365-2141.2005.05409.x. Kubo M, Sonoda Y, Muramatsu R, Usui M. Immunogenicity of human amniotic membrane in experimental xenotransplantation. Invest Ophthalmol Vis Sci. 2001;42(7):1539–46. Friedenstein AJ, Chailakhjan RK, Lalykina KS. The development of fibroblast colonies in monolayer cultures of guinea-pig bone marrow and spleen cells. Cell Tissue Kinet. 1970;3(4):393–403. doi:10.1111/j.1365-2184.1970.tb00347.x. Aust L, Devlin B, Foster SJ, Halvorsen YD, Hicok K, du Laney T, et al. Yield of human adipose-derived adult stem cells from liposuction aspirates. Cytotherapy. 2004;6(1):7–14. doi:10.1080/14653240310004539. Cawthorn WP, Scheller EL, MacDougald OA. Adipose tissue stem cells meet preadipocyte commitment: going back to the future. J Lipid Res. 2012;53(2):227–46. doi:10.1194/jlr.R021089. Cousin B, Andre M, Arnaud E, Penicaud L, Casteilla L. Reconstitution of lethally irradiated mice by cells isolated from adipose tissue. Biochem Biophys Res Commun. 2003;301(4):1016–22. doi:10.1016/S0006-291X(03)00061-5. Han J, Koh YJ, Moon HR, Ryoo HG, Cho CH, Kim I, et al. Adipose tissue is an extramedullary reservoir for functional hematopoietic stem and progenitor cells. Blood. 2010;115(5):957–64. doi:10.1182/blood-2009-05-219923. McIntosh K, Zvonic S, Garrett S, Mitchell JB, Floyd ZE, Hammill L, et al. The immunogenicity of human adipose-derived cells: temporal changes in vitro. Stem Cells. 2006;24(5):1246–53. doi:10.1634/stemcells.2005-0235. Zimmerlin L, Donnenberg VS, Pfeifer ME, Meyer EM, Peault B, Rubin JP, et al. Stromal vascular progenitors in adult human adipose tissue. Cytometry Part A: J Int Soc Anal Cytol. 2010;77(1):22–30. doi:10.1002/cyto.a.20813. Yoshimura K, Asano Y, Aoi N, Kurita M, Oshima Y, Sato K, et al. Progenitor-enriched adipose tissue transplantation as rescue for breast implant complications. Breast J. 2010;16(2):169–75. doi:10.1111/j.1524-4741.2009.00873.x. Sterodimas A, de Faria J, Nicaretta B, Boriani F. Autologous fat transplantation versus adipose-derived stem cell-enriched lipografts: a study. Aesthet Surg J. 2011;31(6):682–93. doi:10.1177/1090820x11415976. Yoshimura K, Sato K, Aoi N, Kurita M, Hirohi T, Harii K. Cell-assisted lipotransfer for cosmetic breast augmentation: supportive use of adipose-derived stem/stromal cells. Aesthetic Plast Surg. 2008;32(1):48–55. doi:10.1007/s00266-007-9019-4. Rigotti G, Marchi A, Galie M, Baroni G, Benati D, Krampera M, et al. Clinical treatment of radiotherapy tissue damage by lipoaspirate transplant: a healing process mediated by adipose-derived adult stem cells. Plast Reconstr Surg. 2007;119(5):1409–22. doi:10.1097/01.prs.0000256047.47909.71. Riordan NH, Ichim TE, Min WP, Wang H, Solano F, Lara F, et al. Non-expanded adipose stromal vascular fraction cell therapy for multiple sclerosis. J Transl Med. 2009;7:29. doi:10.1186/1479-5876-7-29. Michalek J, Moster R, Lukac L, Proefrock K, Petrasovic M, Rybar J, et al. Autologous adipose tissue-derived stromal vascular fraction cells application in patients with osteoarthritis. Cell Transplant. 2015. doi:10.3727/096368915x686760. Bourin P, Bunnell BA, Casteilla L, Dominici M, Katz AJ, March KL, et al. Stromal cells from the adipose tissue-derived stromal vascular fraction and culture expanded adipose tissue-derived stromal/stem cells: a joint statement of the International Federation for Adipose Therapeutics and Science (IFATS) and the International Society for Cellular Therapy (ISCT). Cytotherapy. 2013;15(6):641–8. doi:10.1016/j.jcyt.2013.02.006. Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H, et al. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell. 2002;13(12):4279–95. doi:10.1091/mbc.E02-02-0105. Gimble J, Guilak F. Adipose-derived adult stem cells: isolation, characterization, and differentiation potential. Cytotherapy. 2003;5(5):362–9. doi:10.1080/14653240310003026. Ashjian PH, Elbarbary AS, Edmonds B, DeUgarte D, Zhu M, Zuk PA, et al. In vitro differentiation of human processed lipoaspirate cells into early neural progenitors. Plast Reconstr Surg. 2003;111(6):1922–31. doi:10.1097/01.prs.0000055043.62589.05. Banerjee A, Nurnberger S, Hennerbichler S, Riedl S, Schuh CM, Hacobian A, et al. In toto differentiation of human amniotic membrane towards the Schwann cell lineage. Cell Tissue Bank. 2014;15(2):227–39. doi:10.1007/s10561-013-9401-1. Boquest AC, Shahdadfar A, Fronsdal K, Sigurjonsson O, Tunheim SH, Collas P, et al. Isolation and transcription profiling of purified uncultured human stromal stem cells: alteration of gene expression after in vitro cell culture. Mol Biol Cell. 2005;16(3):1131–41. doi:10.1091/mbc.E04-10-0949. Safford KM, Hicok KC, Safford SD, Halvorsen YD, Wilkison WO, Gimble JM, et al. Neurogenic differentiation of murine and human adipose-derived stromal cells. Biochem Biophys Res Commun. 2002;294(2):371–9. doi:10.1016/s0006-291x(02)00469-2. Planat-Benard V, Menard C, Andre M, Puceat M, Perez A, Garcia-Verdugo JM, et al. Spontaneous cardiomyocyte differentiation from adipose tissue stroma cells. Circ Res. 2004;94(2):223–9. doi:10.1161/01.res.0000109792.43271.47. Rangappa S, Fen C, Lee EH, Bongso A, Sim EK. Transformation of adult mesenchymal stem cells isolated from the fatty tissue into cardiomyocytes. Ann Thorac Surg. 2003;75(3):775–9. Miranville A, Heeschen C, Sengenes C, Curat CA, Busse R, Bouloumie A. Improvement of postnatal neovascularization by human adipose tissue-derived stem cells. Circulation. 2004;110(3):349–55. doi:10.1161/01.cir.0000135466.16823.d0. Planat-Benard V, Silvestre JS, Cousin B, Andre M, Nibbelink M, Tamarat R, et al. Plasticity of human adipose lineage cells toward endothelial cells: physiological and therapeutic perspectives. Circulation. 2004;109(5):656–63. doi:10.1161/01.cir.0000114522.38265.61. Rehman J, Traktuev D, Li J, Merfeld-Clauss S, Temm-Grove CJ, Bovenkerk JE, et al. Secretion of angiogenic and antiapoptotic factors by human adipose stromal cells. Circulation. 2004;109(10):1292–8. doi:10.1161/01.cir.0000121425.42966.f1. Al Battah F, De Kock J, Vanhaecke T, Rogiers V. Current status of human adipose-derived stem cells: differentiation into hepatocyte-like cells. TheScientificWorldJOURNAL. 2011;11:1568–81. doi:10.1100/tsw.2011.146. Seo MJ, Suh SY, Bae YC, Jung JS. Differentiation of human adipose stromal cells into hepatic lineage in vitro and in vivo. Biochem Biophys Res Commun. 2005;328(1):258–64. doi:10.1016/j.bbrc.2004.12.158. Baer PC. Adipose-derived stem cells and their potential to differentiate into the epithelial lineage. Stem Cells Dev. 2011;20(10):1805–16. doi:10.1089/scd.2011.0086. Brzoska M, Geiger H, Gauer S, Baer P. Epithelial differentiation of human adipose tissue-derived adult stem cells. Biochem Biophys Res Commun. 2005;330(1):142–50. doi:10.1016/j.bbrc.2005.02.141. Vossmerbaeumer U, Ohnesorge S, Kuehl S, Haapalahti M, Kluter H, Jonas JB, et al. Retinal pigment epithelial phenotype induced in human adipose tissue-derived mesenchymal stromal cells. Cytotherapy. 2009;11(2):177–88. doi:10.1080/14653240802714819. Du Y, Roh DS, Funderburgh ML, Mann MM, Marra KG, Rubin JP, et al. Adipose-derived stem cells differentiate to keratocytes in vitro. Mol Vis. 2010;16:2680–9. Ferro F, Spelat R, Falini G, Gallelli A, D’Aurizio F, Puppato E, et al. Adipose tissue-derived stem cell in vitro differentiation in a three-dimensional dental bud structure. Am J Pathol. 2011;178(5):2299–310. doi:10.1016/j.ajpath.2011.01.055. 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(2):103–10. doi:10.2174/157488810791268564. Tobita M, Orbay H, Mizuno H. Adipose-derived stem cells: current findings and future perspectives. Discov Med. 2011;11(57):160–70. Frazier TP, Gimble JM, Kheterpal I, Rowan BG. Impact of low oxygen on the secretome of human adipose-derived stromal/stem cell primary cultures. Biochimie. 2013;95(12):2286–96. doi:10.1016/j.biochi.2013.07.011. Moon KM, Park YH, Lee JS, Chae YB, Kim MM, Kim DS, et al. The effect of secretory factors of adipose-derived stem cells on human keratinocytes. Int J Mol Sci. 2012;13(1):1239–57. doi:10.3390/ijms13011239. Kutten JC, McGovern D, Hobson CM, Luffy SA, Nieponice A, Tobita K, et al. Decellularized tracheal extracellular matrix supports epithelial migration, differentiation, and function. Tissue Eng A. 2015;21(1–2):75–84. doi:10.1089/ten.TEA.2014.0089. Hassan WU, Greiser U, Wang W. Role of adipose-derived stem cells in wound healing. Wound repair and regeneration: official publication of the Wound Healing Society [and] the European Tissue Repair. Society. 2014;22(3):313–25. doi:10.1111/wrr.12173. Kilroy GE, Foster SJ, Wu X, Ruiz J, Sherwood S, Heifetz A, et al. Cytokine profile of human adipose-derived stem cells: expression of angiogenic, hematopoietic, and pro-inflammatory factors. J Cell Physiol. 2007;212(3):702–9. doi:10.1002/jcp.21068. Kronsteiner B, Peterbauer-Scherb A, Grillari-Voglauer R, Redl H, Gabriel C, van Griensven M, et al. Human mesenchymal stem cells and renal tubular epithelial cells differentially influence monocyte-derived dendritic cell differentiation and maturation. Cell Immunol. 2011;267(1):30–8. doi:10.1016/j.cellimm.2010.11.001. Wolbank S, Peterbauer A, Fahrner M, Hennerbichler S, van Griensven M, Stadler G, et al. Dose-dependent immunomodulatory effect of human stem cells from amniotic membrane: a comparison with human mesenchymal stem cells from adipose tissue. Tissue Eng. 2007;13(6):1173–83. doi:10.1089/ten.2006.0313. Wolbank S, Stadler G, Peterbauer A, Gillich A, Karbiener M, Streubel B, et al. Telomerase immortalized human amnion- and adipose-derived mesenchymal stem cells: maintenance of differentiation and immunomodulatory characteristics. Tissue Eng A. 2009;15(7):1843–54. doi:10.1089/ten.tea.2008.0205. Kronsteiner B, Wolbank S, Peterbauer A, Hackl C, Redl H, van Griensven M, et al. Human mesenchymal stem cells from adipose tissue and amnion influence T-cells depending on stimulation method and presence of other immune cells. Stem Cells Dev. 2011;20(12):2115–26. doi:10.1089/scd.2011.0031. Cho YB, Park KJ, Yoon SN, Song KH, Kim DS, Jung SH et al. Long-term results of adipose-derived stem cell therapy for the treatment of Crohn’s fistula. Stem Cells Transl Med. 2015. doi:10.5966/sctm.2014-0199. Gir P, Oni G, Brown SA, Mojallal A, Rohrich RJ. Human adipose stem cells: current clinical applications. Plast Reconstr Surg. 2012;129(6):1277–90. doi:10.1097/PRS.0b013e31824ecae6. Mizuno H, Tobita M, Uysal AC. Concise review: adipose-derived stem cells as a novel tool for future regenerative medicine. Stem Cells. 2012;30(5):804–10. doi:10.1002/stem.1076. Gimble JM, Guilak F, Bunnell BA. Clinical and preclinical translation of cell-based therapies using adipose tissue-derived cells. Stem Cell Res Ther. 2010;1(2):19. doi:10.1186/scrt19. Sergeevicheva V, Kruchkova I, Chernykh E, Shevela E, Kulagin A, Gilevich A, et al. Rapid recovery from chronic PRCA by MSC infusion in patient after major ABO-mismatched alloSCT. Case Rep Med. 2012;2012:862721. doi:10.1155/2012/862721. Ra JC, Kang SK, Shin IS, Park HG, Joo SA, Kim JG, et al. Stem cell treatment for patients with autoimmune disease by systemic infusion of culture-expanded autologous adipose tissue derived mesenchymal stem cells. J Transl Med. 2011;9:181. doi:10.1186/1479-5876-9-181. Bura A, Planat-Benard V, Bourin P, Silvestre JS, Gross F, Grolleau JL, et al. Phase I trial: the use of autologous cultured adipose-derived stroma/stem cells to treat patients with non-revascularizable critical limb ischemia. Cytotherapy. 2014;16(2):245–57. doi:10.1016/j.jcyt.2013.11.011. Sandor GK, Numminen J, Wolff J, Thesleff T, Miettinen A, Tuovinen VJ, et al. Adipose stem cells used to reconstruct 13 cases with cranio-maxillofacial hard-tissue defects. Stem Cells Transl Med. 2014;3(4):530–40. doi:10.5966/sctm.2013-0173. Thesleff T, Lehtimaki K, Niskakangas T, Mannerstrom B, Miettinen S, Suuronen R, et al. Cranioplasty with adipose-derived stem cells and biomaterial: a novel method for cranial reconstruction. Neurosurgery. 2011;68(6):1535–40. doi:10.1227/NEU.0b013e31820ee24e. Tanikawa DY, Aguena M, Bueno DF, Passos-Bueno MR, Alonso N. Fat grafts supplemented with adipose-derived stromal cells in the rehabilitation of patients with craniofacial microsomia. Plast Reconstr Surg. 2013;132(1):141–52. doi:10.1097/PRS.0b013e3182910a82. Zuk P. Adipose-derived stem cells in tissue regeneration: a review. ISRN Stem Cells. 2013;2013:35. doi:10.1155/2013/713959. Castro-Malaspina H, Gay RE, Resnick G, Kapoor N, Meyers P, Chiarieri D, et al. Characterization of human bone marrow fibroblast colony-forming cells (CFU-F) and their progeny. Blood. 1980;56(2):289–301. EudraLex. Clinical trial guidelines. 2010 (Volume 10). EudraLex. Good manufacturing practice (GMP). 2015 (Volume 4). Aarya Hari SG. Production of good manufacturing practice grade equine adiposederived mesenchymal stem cells for therapeutic use. J Stem Cell Res Ther. 2013;03(05):2157–7633. doi:10.4172/2157-7633.1000154. Sensebe L, Gadelorge M, Fleury-Cappellesso S. Production of mesenchymal stromal/stem cells according to good manufacturing practices: a review. Stem Cell Res Ther. 2013;4(3):66. doi:10.1186/scrt217. Carvalho PP, Gimble JM, Dias IR, Gomes ME, Reis RL. Xenofree enzymatic products for the isolation of human adipose-derived stromal/stem cells. Tissue Eng Part C Meth. 2013;19(6):473–8. doi:10.1089/ten.TEC.2012.0465. Patrikoski M, Juntunen M, Boucher S, Campbell A, Vemuri MC, Mannerstrom B, et al. Development of fully defined xeno-free culture system for the preparation and propagation of cell therapy-compliant human adipose stem cells. Stem Cell Res Ther. 2013;4(2):27. doi:10.1186/scrt175. Aguena M, Fanganiello RD, Tissiani LA, Ishiy FA, Atique R, Alonso N, et al. Optimization of parameters for a more efficient use of adipose-derived stem cells in regenerative medicine therapies. Stem Cells Int. 2012;2012:303610. doi:10.1155/2012/303610. Yang XF, He X, He J, Zhang LH, Su XJ, Dong ZY, et al. High efficient isolation and systematic identification of human adipose-derived mesenchymal stem cells. J Biomed Sci. 2011;18:59. doi:10.1186/1423-0127-18-59. Markarian CF, Frey GZ, Silveira MD, Chem EM, Milani AR, Ely PB, et al. Isolation of adipose-derived stem cells: a comparison among different methods. Biotechnol Lett. 2014;36(4):693–702. doi:10.1007/s10529-013-1425-x. Philips BJ, Marra KG, Rubin JP. Adipose stem cell-based soft tissue regeneration. Expert Opin Biol Ther. 2012;12(2):155–63. doi:10.1517/14712598.2012.644533. Thirumala S, Gimble JM, Devireddy RV. Cryopreservation of stromal vascular fraction of adipose tissue in a serum-free freezing medium. J Tissue Eng Regen Med. 2010;4(3):224–32. doi:10.1002/term.232. Guven S, Karagianni M, Schwalbe M, Schreiner S, Farhadi J, Bula S, et al. Validation of an automated procedure to isolate human adipose tissue-derived cells by using the Sepax(R) technology. Tissue Eng Part C Methods. 2012;18(8):575–82. doi:10.1089/ten.TEC.2011.0617. Lin K, Matsubara Y, Masuda Y, Togashi K, Ohno T, Tamura T, et al. Characterization of adipose tissue-derived cells isolated with the Celution system. Cytotherapy. 2008;10(4):417–26. doi:10.1080/14653240801982979. Scherberich A, Galli R, Jaquiery C, Farhadi J, Martin I. Three-dimensional perfusion culture of human adipose tissue-derived endothelial and osteoblastic progenitors generates osteogenic constructs with intrinsic vascularization capacity. Stem Cells. 2007;25(7):1823–9. doi:10.1634/stemcells.2007-0124. Lin G, Garcia M, Ning H, Banie L, Guo YL, Lue TF, et al. Defining stem and progenitor cells within adipose tissue. Stem Cells Dev. 2008;17(6):1053–63. doi:10.1089/scd.2008.0117. Eom YW, Lee JE, Yang MS, Jang IK, Kim HE, Lee DH, et al. Rapid isolation of adipose tissue-derived stem cells by the storage of lipoaspirates. Yonsei Med J. 2011;52(6):999–1007. doi:10.3349/ymj.2011.52.6.999. Suga H, Shigeura T, Matsumoto D, Inoue K, Kato H, Aoi N, et al. Rapid expansion of human adipose-derived stromal cells preserving multipotency. Cytotherapy. 2007;9(8):738–45. doi:10.1080/14653240701679873. Safwani WK, Makpol S, Sathapan S, Chua KH. Alteration of gene expression levels during osteogenic induction of human adipose derived stem cells in long-term culture. Cell Tissue Bank. 2013;14(2):289–301. doi:10.1007/s10561-012-9309-1. Kirkpatrick CJ, Melzner I, Goller T. Comparative effects of trypsin, collagenase and mechanical harvesting on cell membrane lipids studied in monolayer-cultured endothelial cells and a green monkey kidney cell line. Biochim Biophys Acta. 1985;846(1):120–6. Stadler G, Hennerbichler S, Lindenmair A, Peterbauer A, Hofer K, van Griensven M, et al. Phenotypic shift of human amniotic epithelial cells in culture is associated with reduced osteogenic differentiation in vitro. Cytotherapy. 2008;10(7):743–52. doi:10.1080/14653240802345804. Kakagia D, Pallua N. Autologous fat grafting: in search of the optimal technique. Surg Innov. 2014;21(3):327–36. doi:10.1177/1553350613518846. Garza RM, Paik KJ, Chung MT, Duscher D, Gurtner GC, Longaker MT, et al. Studies in fat grafting: part III. Fat grafting irradiated tissue—improved skin quality and decreased fat graft retention. Plast Reconstr Surg. 2014;134(2):249–57. doi:10.1097/prs.0000000000000326. Stillaert FB, Di Bartolo C, Hunt JA, Rhodes NP, Tognana E, Monstrey S, et al. Human clinical experience with adipose precursor cells seeded on hyaluronic acid-based spongy scaffolds. Biomaterials. 2008;29(29):3953–9. doi:10.1016/j.biomaterials.2008.06.005. Matsumoto D, Sato K, Gonda K, Takaki Y, Shigeura T, Sato T, et al. Cell-assisted lipotransfer: supportive use of human adipose-derived cells for soft tissue augmentation with lipoinjection. Tissue Eng. 2006;12(12):3375–82. doi:10.1089/ten.2006.12.3375. Holnthoner W, Hohenegger K, Husa AM, Muehleder S, Meinl A, Peterbauer-Scherb A, et al. Adipose-derived stem cells induce vascular tube formation of outgrowth endothelial cells in a fibrin matrix. J Tissue Eng Regen Med. 2015;9(2):127–36. doi:10.1002/term.1620. Gentile P, Orlandi A, Scioli MG, Di Pasquali C, Bocchini I, Curcio CB, et al. A comparative translational study: the combined use of enhanced stromal vascular fraction and platelet-rich plasma improves fat grafting maintenance in breast reconstruction. Stem Cells Transl Med. 2012;1(4):341–51. doi:10.5966/sctm.2011-0065. Jiang A, Li M, Duan W, Dong Y, Wang Y. Improvement of the survival of human autologous fat transplantation by adipose-derived stem-cells-assisted lipotransfer combined with bFGF. TheScientificWorldJOURNAL. 2015;2015:968057. doi:10.1155/2015/968057. Luo S, Hao L, Li X, Yu D, Diao Z, Ren L, et al. Adipose tissue-derived stem cells treated with estradiol enhance survival of autologous fat transplants. Tohoku J Exp Med. 2013;231(2):101–10. Li L, Pan S, Ni B, Lin Y. Improvement in autologous human fat transplant survival with SVF plus VEGF-PLA nano-sustained release microspheres. Cell Biol Int. 2014;38(8):962–70. doi:10.1002/cbin.10284. Van Pham P, Hong-Thien Bui K, Quoc Ngo D, Tan Khuat L, Kim PN. Transplantation of nonexpanded adipose stromal vascular fraction and platelet-rich plasma for articular cartilage injury treatment in mice model. J Med Eng. 2013;2013:7. doi:10.1155/2013/832396. Paspaliaris B, Thornton JAF. Methods and apparatuses for isolating and preparing stem cells. 2014. US 20140093482 A1. Tzouvelekis A, Koliakos G, Ntolios P, Baira I, Bouros E, Oikonomou A, et al. Stem cell therapy for idiopathic pulmonary fibrosis: a protocol proposal. J Transl Med. 2011;9:182. doi:10.1186/1479-5876-9-182. Tzouvelekis A, Paspaliaris V, Koliakos G, Ntolios P, Bouros E, Oikonomou A, et al. A prospective, non-randomized, no placebo-controlled, phase Ib clinical trial to study the safety of the adipose derived stromal cells-stromal vascular fraction in idiopathic pulmonary fibrosis. J Transl Med. 2013;11:171. doi:10.1186/1479-5876-11-171. Aronowitz JA, Ellenhorn JD. Adipose stromal vascular fraction isolation: a head-to-head comparison of four commercial cell separation systems. Plast Reconstr Surg. 2013;132(6):932e–9. doi:10.1097/PRS.0b013e3182a80652. Kakudo N, Tanaka Y, Morimoto N, Ogawa T, Kushida S, Hara T, et al. Adipose-derived regenerative cell (ADRC)-enriched fat grafting: optimal cell concentration and effects on grafted fat characteristics. J Transl Med. 2013;11:254. doi:10.1186/1479-5876-11-254. Granel B, Daumas A, Jouve E, Harle JR, Nguyen PS, Chabannon C, et al. Safety, tolerability and potential efficacy of injection of autologous adipose-derived stromal vascular fraction in the fingers of patients with systemic sclerosis: an open-label phase I trial. Ann Rheum Dis. 2014. doi:10.1136/annrheumdis-2014-205681. Gotoh M, Yamamoto T, Kato M, Majima T, Toriyama K, Kamei Y, et al. Regenerative treatment of male stress urinary incontinence by periurethral injection of autologous adipose-derived regenerative cells: 1-year outcomes in 11 patients. Int J Urol. 2014;21(3):294–300. doi:10.1111/iju.12266. Perez-Cano R, Vranckx JJ, Lasso JM, Calabrese C, Merck B, Milstein AM, et al. Prospective trial of adipose-derived regenerative cell (ADRC)-enriched fat grafting for partial mastectomy defects: the RESTORE-2 trial. Eur J Surg Oncol. 2012;38(5):382–9. doi:10.1016/j.ejso.2012.02.178. Dos-Anjos Vilaboa S, Navarro-Palou M, Llull R. Age influence on stromal vascular fraction cell yield obtained from human lipoaspirates. Cytotherapy. 2014;16(8):1092–7. doi:10.1016/j.jcyt.2014.02.007. Pak J, Lee JH, Lee SH. Regenerative repair of damaged meniscus with autologous adipose tissue-derived stem cells. BioMed Res Int. 2014;2014:436029. doi:10.1155/2014/436029. Pak J, Chang JJ, Lee JH, Lee SH. Safety reporting on implantation of autologous adipose tissue-derived stem cells with platelet-rich plasma into human articular joints. BMC Musculoskelet Disord. 2013;14:337. doi:10.1186/1471-2474-14-337. Khan ZA, Dulgar-Tulloch AJ, Rakuff S, Shoemaker PA, Kvam EL, Chen X, et al. Automated systems and methods for isolating regenerative cells from adipose tissue. 2012. US 20120276628 A1. Kaengkan P, Baek SE, Kim JY, Kam KY, Do BR, Lee ES, et al. Administration of mesenchymal stem cells and ziprasidone enhanced amelioration of ischemic brain damage in rats. Mol Cells. 2013;36(6):534–41. doi:10.1007/s10059-013-0235-2. Stubbers R, Coleman ME. Apparatus and methods for cell isolation. 2015. US 20150056691 A1. Doi K, Tanaka S, Iida H, Eto H, Kato H, Aoi N, et al. Stromal vascular fraction isolated from lipo-aspirates using an automated processing system: bench and bed analysis. J Tissue Eng Regen Med. 2013;7(11):864–70. doi:10.1002/term.1478. Domenis R, Lazzaro L, Calabrese S, Mangoni D, Gallelli A, Bourkoula E, et al. Adipose tissue derived stem cells: in vitro and in vivo analysis of a standard and three commercially available cell-assisted lipotransfer techniques. Stem Cell Res Ther. 2015;6(1):2. doi:10.1186/scrt536. Schafer ME, Hicok KC, Mills DC, Cohen SR, Chao JJ. Acute adipocyte viability after third-generation ultrasound-assisted liposuction. Aesthet Surg J Am Soc Aesthet Plast Surg. 2013;33(5):698–704. doi:10.1177/1090820x13485239. Zhu M, Cohen SR, Hicok KC, Shanahan RK, Strem BM, Yu JC, et al. Comparison of three different fat graft preparation methods: gravity separation, centrifugation, and simultaneous washing with filtration in a closed system. Plast Reconstr Surg. 2013;131(4):873–80. doi:10.1097/PRS.0b013e31828276e9. Ansorge H, Garza JR, McCormack MC, Leamy P, Roesch S, Barere A, et al. Autologous fat processing via the Revolve system: quality and quantity of fat retention evaluated in an animal model. Aesthet Surg J Am Soc Aesthet Plast Sur. 2014;34(3):438–47. doi:10.1177/1090820x14524416. Dos-Anjos Vilaboa S, Llull R, Mendel TA. Returning fat grafts to physiologic conditions using washing. Plast Reconstr Surg. 2013;132(2):323e–6. doi:10.1097/PRS.0b013e3182958be1. Zimmerlin L, Rubin JP, Pfeifer ME, Moore LR, Donnenberg VS, Donnenberg AD. Human adipose stromal vascular cell delivery in a fibrin spray. Cytotherapy. 2013;15(1):102–8. doi:10.1016/j.jcyt.2012.10.009. Ferguson RE, Cui X, Fink BF, Vasconez HC, Pu LL. The viability of autologous fat grafts harvested with the LipiVage system: a comparative study. Ann Plast Surg. 2008;60(5):594–7. doi:10.1097/SAP.0b013e31817433c5. Bianchi F, Maioli M, Leonardi E, Olivi E, Pasquinelli G, Valente S, et al. A new nonenzymatic method and device to obtain a fat tissue derivative highly enriched in pericyte-like elements by mild mechanical forces from human lipoaspirates. Cell Transplant. 2013;22(11):2063–77. doi:10.3727/096368912X657855. García-Contreras M, Messaggio F, Jimenez O, Mendez A. Differences in exosome content of human adipose tissue processed by non-enzymatic and enzymatic methods. CellR4. 2015;3(1):e1423. Bianchi F, Olivi E, Baldassarre M, Giannone FA, Laggetta M, Valente S, et al. Lipogems, a new modality of fat tissue handling to enhance tissue repair in chronic hind limb ischemia. CellR4. 2014;2(6):e1289. Maioli M, Rinaldi S, Santaniello S, Castagna A, Pigliaru G, Delitala A, et al. Radioelectric asymmetric conveyed fields and human adipose-derived stem cells obtained with a nonenzymatic method and device: a novel approach to multipotency. Cell Transplant. 2014;23(12):1489–500. doi:10.3727/096368913x672037. Carelli S, Messaggio F, Canazza A, Hebda DM, Caremoli F, Latorre E, et al. Characteristics and properties of mesenchymal stem cells derived from micro-fragmented adipose tissue. Cell Transplant. 2014. doi:10.3727/096368914x681603. Raffaini M, Tremolada C. Micro fractured and purified adipose tissue graft (Lipogems®) can improve the orthognathic surgery outcomes both aesthetically and in postoperative healing. CellR4. 2014;2(0):e1118. Benzi R, Marfia G, Bosetti M, Beltrami G, Magri AS, Versari S et al. Microfractured lipoaspirate may help oral bone and soft tissue regeneration: a case report. CellR4. 2015;3(3):e1583. Giori A, Tremolada C, Vailati R, Navone SE, Marfia G, Caplan AI. Recovery of function in anal incontinence after micro-fragmented fat graft (Lipogems®) injection: two years follow up of the first 5 cases. CellR4. 2015;3(2):e1544. Hu CB, Myers KE, Peterson RC. Devices for harvesting and homogenizing adipose tissue containing autologous endothelial cells. 2000. US 6020196 A1. Victor S. Isolation of stromal vascular fraction from adipose tissue obtained from postmortem source using ultrasonic cavitation. 2014. WO 2014015229 A1. Bright R, Bright P, Hansen B, Thomas W. Isolation of stem cells from adipose tissue by ultrasonic cavitation, and methods of use. 2014. WO 2014000031 A1. Bright R, Bright M, Bright P, Hayne S, Thomas WD. Migraine and tension-type headache treated with stromal vascular fraction: a case series. J Med Case Rep. 2014;8:237. doi:10.1186/1752-1947-8-237. Schafer ME. Selective lysing of cells using ultrasound. 2013. US 20130012927 A1. Gimble JM, Shah FS, Wu X. Non-enzymatic method for isolating human adipose-derived stromal stem cells. 2014. US 20140017783 A1. Michalek J. Method for isolation of adipose tissue-derived stromal vascular fraction cells. 2014. WO 2014169885 A1. Hicok KC, Hedrick MH. Automated isolation and processing of adipose-derived stem and regenerative cells. Methods Mol Biol (Clifton, NJ). 2011;702:87–105. doi:10.1007/978-1-61737-960-4_8. Peterson A, Fornace L. Systems, methods and compositions for optimizing tissue and cell enriched grafts. 2010. US 20100279405 A1. Hedrick MH, Fraser JK, Schulzki MJ, Byrnes B, Carlson G, Schreiber RE, et al. Systems and methods for isolating and using clinically safe adipose derived regenerative cells. 2012. US 20120264200 A1. Kakudo N, Morimoto N, Ogawa T, Kusumoto K. Potential of adipose-derived stem cells for regeneration medicine: clinical application and usefulness of fat grafting. J Stem Cell Res Ther. 2014;4(204):2. Sanchez PL, Sanz-Ruiz R, Fernandez-Santos ME, Fernandez-Aviles F. Cultured and freshly isolated adipose tissue-derived cells: fat years for cardiac stem cell therapy. Eur Heart J. 2010;31(4):394–7. doi:10.1093/eurheartj/ehp403. Cimino WW, Llull R, Katz AJ. Tissue processing apparatus and method for processing adipose tissue. 2015. WO 2015035221 A1. Llull R, Katz AJ, Cimino WW. Method for processing adipose tissue and processing apparatus. 2013. WO 2013106655 A1. Do BR, Lee JK, Kim JH, Pak SH, Shin BS. Peristaltic pump, and regenerative cell extraction system using same. 2013. WO 2013089481 A1. Raj SS, Gopal V, Priya N, Krishnegowda B, Thiruvampattil P, Majumdar AS, et al. System for isolating stromal vascular fraction (svf) cells from the adipose tissue and a method thereof. 2015. US 20150004702 A1. Williams SK, Kosnik P, England C, Cannon TF, Vossman E, Boland E, et al. Apparatus and methods for preparing tissue grafts. 2007. WO 2007009036 A9. Wolters R, Yang A, NELSON J, Williams SK, et al. Hand-held micro-liposuction adipose harvester, processor, and cell concentrator. 2014. WO 2014047368 A1. Ariff GD, Cannon T, Case JL, Haller CL, Kosnik P, Luddy CP, et al. Cell separation apparatus and methods of use. 2008. WO 2008133874 A1. SHIGEKI S, KIAT OW. Method for isolating stromal vascular fraction. 2015. WO 2015005871 A1. Buss B. Autologous tissue harvesting and irrigation device. 2012. US 8202493 A1. Kensy A, Winkler KW. Procedure and device for separating adult stem cells from fatty tissue. 2014. US 8628950 A1. Winkler KW, Matthiesen ID. Device for separating adult stem cells. 2013. EP 2677024 A1. Matthiesen I, Winkler KW. Vorrichtung zum Separieren von adulten Stammzellen. 2013. DE 102013209718 A1. Victor S. Ultrasonic cavitation derived stromal or mesenchymal vascular extracts and cells derived therefrom obtained from adipose tissue and use thereof. 2013. US 20130189234 A1. Victor S. Isolation of stromal vascular fraction from vascular tissues. 2014. WO 2014138383 A1. Cimino WW, Katz AJ, Llull R. Apparatus and methods relating to collecting and processing human biological material containing adipose. 2012. WO 2012006587 A2. Vossman E, Iwami S, Yang A, Cannon T, Paek HJ. Adipose tissue collection and pre-processing devices for use in liposuction procedure. 2014. US 20140193852 A9. Tremolada C. Device and method for preparing tissue, particularly adipose tissue. 2013. US 20130123747 A1. Chapman JR, Sparks R. Apparatus for centrifugation and methods therefore. 2013. WO 2013122683 A1. Shah FS, Wu X, Dietrich M, Rood J, Gimble JM. A non-enzymatic method for isolating human adipose tissue-derived stromal stem cells. Cytotherapy. 2013;15(8):979–85. doi:10.1016/j.jcyt.2013.04.001.