Characterization of adipose tissue-derived stromal vascular fraction for clinical application to cartilage regeneration

Abstract

Bone marrow concentration (BMC) is the most recognized procedure to prepare mesenchymal stem cells for cartilage regeneration. However, bone marrow aspiration is highly invasive and results in low stem cell numbers. Recently, adipose tissue-derived stromal vascular fraction (AT-SVF) was studied as an alternate source of stem cells for cartilage regeneration. However, AT-SVF is not fully characterized in terms of functional equivalence to BMC. Therefore, in this study, we characterized AT-SVF and assessed its suitability as a one-step surgical procedure for cartilage regeneration, as an alternative to BMC. AT-SVF contained approximately sixfold less nucleated cells than BMC. However, adherent cells in AT-SVF were fourfold greater than BMC. Additionally, the colony-forming unit frequency of AT-SVF was higher than that of BMC, at 0.5 and 0.01%, respectively. The mesenchymal stem cell (MSC) population (CD45−CD31−CD90+CD105+) was 4.28% in AT-SVF and 0.42% in BMC, and the adipose-derived stromal cell (ASC) population (CD34+CD31−CD146−) was 32% in AT-SVF and 0.16% in BMC. In vitro chondrogenesis demonstrated that micromass was not formed in BMC, whereas it was clearly formed in AT-SVF. Taken together, uncultured AT-SVF could be used in one-step surgery for cartilage regeneration as a substitute for BMC.

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References

  1. Atalay S, Coruh A, Deniz K (2014) Stromal vascular fraction improves deep partial thickness burn wound healing. Burns: J Int Soc Burn Inj. doi:10.1016/j.burns.2014.01.023

    Google Scholar 

  2. Bartha L, Vajda A, Duska Z, Rahmeh H, Hangody L (2006) Autologous osteochondral mosaicplasty grafting. J Orthop Sports Phys Ther 36:739–750

    Article  PubMed  Google Scholar 

  3. Bartlett W, Skinner JA, Gooding CR, Carrington RW, Flanagan AM, Briggs TW, Bentley G (2005) Autologous chondrocyte implantation versus matrix-induced autologous chondrocyte implantation for osteochondral defects of the knee: a prospective, randomised study. J Bone Jt Surg Brit Vol 87:640–645

    Article  CAS  Google Scholar 

  4. Bourin P, Bunnell BA, Casteilla L, Dominici M, Katz AJ, March KL, Redl H, Rubin JP, Yoshimura K, Gimble JM (2013) 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 15:641–648

    Article  PubMed Central  PubMed  Google Scholar 

  5. Brittberg M (1999) Autologous chondrocyte transplantation. Clin Ortho Relat Res: S147–155

  6. Brittberg M, Lindahl A, Nilsson A, Ohlsson C, Isaksson O, Peterson L (1994) Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation. N Engl J Med 331:889–895

    Article  CAS  PubMed  Google Scholar 

  7. Buda R, Vannini F, Cavallo M, Grigolo B, Cenacchi A, Giannini S (2010) Osteochondral lesions of the knee: a new one-step repair technique with bone-marrow-derived cells. J Bone Jt Surg Am Vol 92(Suppl 2):2–11

    Article  Google Scholar 

  8. Cavallo C, Desando G, Columbaro M, Ferrari A, Zini N, Facchini A, Grigolo B (2013) Chondrogenic differentiation of bone marrow concentrate grown onto a hylauronan scaffold: rationale for its use in the treatment of cartilage lesions. J Biomed Mater Res Part A 101:1559–1570

    Article  Google Scholar 

  9. De Ugarte DA, Morizono K, Elbarbary A, Alfonso Z, Zuk PA, Zhu M, Dragoo JL, Ashjian P, Thomas B, Benhaim P, Chen I, Fraser J, Hedrick MH (2003) Comparison of multi-lineage cells from human adipose tissue and bone marrow. Cells Tissues Organs 174:101–109

    Article  PubMed  Google Scholar 

  10. Diaz-Romero J, Gaillard JP, Grogan SP, Nesic D, Trub T, Mainil-Varlet P (2005) Immunophenotypic analysis of human articular chondrocytes: changes in surface markers associated with cell expansion in monolayer culture. J Cell Physiol 202:731–742

    Article  CAS  PubMed  Google Scholar 

  11. English A, Jones EA, Corscadden D, Henshaw K, Chapman T, Emery P, McGonagle D (2007) A comparative assessment of cartilage and joint fat pad as a potential source of cells for autologous therapy development in knee osteoarthritis. Rheumatology 46:1676–1683

    Article  CAS  PubMed  Google Scholar 

  12. Fortier LA, Potter HG, Rickey EJ, Schnabel LV, Foo LF, Chong LR, Stokol T, Cheetham J, Nixon AJ (2010) Concentrated bone marrow aspirate improves full-thickness cartilage repair compared with microfracture in the equine model. J Bone Jt Surg Am Vol 92:1927–1937

    Article  Google Scholar 

  13. Gobbi A, Karnatzikos G, Scotti C, Mahajan V, Mazzucco L, Grigolo B (2011) One-step cartilage repair with bone marrow aspirate concentrated cells and collagen matrix in full-thickness knee cartilage lesions: results at 2-year follow-up. Cartil 2:286–299

    Article  Google Scholar 

  14. Hermann PC, Huber SL, Herrler T, von Hesler C, Andrassy J, Kevy SV, Jacobson MS, Heeschen C (2008) Concentration of bone marrow total nucleated cells by a point-of-care device provides a high yield and preserves their functional activity. Cell Transplant 16:1059–1069

    Article  PubMed  Google Scholar 

  15. Hunziker EB (2002) Articular cartilage repair: basic science and clinical progress. A review of the current status and prospects. Osteoarthritis and cartilage/OARS. Osteoarthr Res Soc 10:432–463

    Article  CAS  Google Scholar 

  16. Jankowski RJ, Deasy BM, Huard J (2002) Muscle-derived stem cells. Gene Ther 9:642–647

    Article  CAS  PubMed  Google Scholar 

  17. Jiang T, Liu W, Lv X, Sun H, Zhang L, Liu Y, Zhang WJ, Cao Y, Zhou G (2010) Potent in vitro chondrogenesis of CD105 enriched human adipose-derived stem cells. Biomaterials 31:3564–3571

  18. Jiang Y, Vaessen B, Lenvik T, Blackstad M, Reyes M, Verfaillie CM (2002) Multipotent progenitor cells can be isolated from postnatal murine bone marrow, muscle, and brain. Exp Hematol 30:896–904

    Article  CAS  PubMed  Google Scholar 

  19. Jurgens WJ, van Dijk A, Doulabi BZ, Niessen FB, Ritt MJ, van Milligen FJ, Helder MN (2009) Freshly isolated stromal cells from the infrapatellar fat pad are suitable for a one-step surgical procedure to regenerate cartilage tissue. Cytotherapy 11:1052–1064

    Article  CAS  PubMed  Google Scholar 

  20. Khan MH (2012) Update on liposuction: clinical pearls. Cutis 90:259–265

    PubMed  Google Scholar 

  21. Klein JA (1990) The tumescent technique. Anesthesia and modified liposuction technique. Dermatol Clin 8:425–437

    CAS  PubMed  Google Scholar 

  22. Koh YG, Choi YJ (2012) Infrapatellar fat pad-derived mesenchymal stem cell therapy for knee osteoarthritis. Knee 19:902–907

    Article  PubMed  Google Scholar 

  23. Koh YG, Choi YJ, Kwon SK, Kim YS, Yeo JE (2013) Clinical results and second-look arthroscopic findings after treatment with adipose-derived stem cells for knee osteoarthritis. Knee Surg Sports Traumatol Arthrosc: Off J ESSKA. doi:10.1007/s00167-013-2807-2

    Google Scholar 

  24. Kokai LE, Marra K, Rubin JP (2013) Adipose stem cells: biology and clinical applications for tissue repair and regeneration. Transl Res: J Lab Clin Med. doi:10.1016/j.trsl.2013.11.009

    Google Scholar 

  25. Leblanc AJ, Touroo JS, Hoying JB, Williams SK (2012) Adipose stromal vascular fraction cell construct sustains coronary microvascular function after acute myocardial infarction. Am J Physiol Heart Circ Physiol 302:H973–H982

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  26. Lechner A, Habener JF (2003) Stem/progenitor cells derived from adult tissues: potential for the treatment of diabetes mellitus. Am J Physiol Endocrinol Metab 284:E259–E266

    CAS  PubMed  Google Scholar 

  27. Martin DR, Cox NR, Hathcock TL, Niemeyer GP, Baker HJ (2002) Isolation and characterization of multipotential mesenchymal stem cells from feline bone marrow. Exp Hematol 30:879–886

    Article  CAS  PubMed  Google Scholar 

  28. 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 24:376–385

    Article  PubMed  Google Scholar 

  29. Murawski CD, Foo LF, Kennedy JG (2010) A review of arthroscopic bone marrow stimulation techniques of the talus: the good, the bad, and the causes for concern. Cartil 1:137–144

    Article  Google Scholar 

  30. Nagase T, Muneta T, Ju YJ, Hara K, Morito T, Koga H, Nimura A, Mochizuki T, Sekiya I (2008) Analysis of the chondrogenic potential of human synovial stem cells according to harvest site and culture parameters in knees with medial compartment osteoarthritis. Arthritis Rheum 58:1389–1398

    Article  PubMed  Google Scholar 

  31. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147

    Article  CAS  PubMed  Google Scholar 

  32. Premaratne GU, Ma LP, Fujita M, Lin X, Bollano E, Fu M (2011) Stromal vascular fraction transplantation as an alternative therapy for ischemic heart failure: anti-inflammatory role. J Cardiothorac Surg 6:43

    Article  PubMed Central  PubMed  Google Scholar 

  33. Sheng L, Yang M, Du Z, Yang Y, Li Q (2013) Transplantation of stromal vascular fraction as an alternative for accelerating tissue expansion. J Plast Reconstr Aesthet Surg: JPRAS 66:551–557

    Article  PubMed  Google Scholar 

  34. Shi W, Wang H, Pan G, Geng Y, Guo Y, Pei D (2006) Regulation of the pluripotency marker Rex-1 by Nanog and Sox2. J Biol Chem 281:23319–23325

    Article  CAS  PubMed  Google Scholar 

  35. Steadman JR, Rodkey WG, Rodrigo JJ (2001) Microfracture: surgical technique and rehabilitation to treat chondral defects. Clin Orthop Relat Res: S362–369

  36. Varma MJ, Breuls RG, Schouten TE, Jurgens WJ, Bontkes HJ, Schuurhuis GJ, van Ham SM, van Milligen FJ (2007) Phenotypical and functional characterization of freshly isolated adipose tissue-derived stem cells. Stem Cells Dev 16:91–104

    Article  PubMed  Google Scholar 

  37. Yoon DS, Kim YH, Jung HS, Paik S, Lee JW (2011) Importance of Sox2 in maintenance of cell proliferation and multipotency of mesenchymal stem cells in low-density culture. Cell Prolif 44:428–440

    Article  CAS  PubMed  Google Scholar 

  38. Yu J, Vodyanik MA, Smuga-Otto K, Antosiewicz-Bourget J, Frane JL, Tian S, Nie J, Jonsdottir GA, Ruotti V, Stewart R, Slukvin II, Thomson JA (2007) Induced pluripotent stem cell lines derived from human somatic cells. Science 318:1917–1920

    Article  CAS  PubMed  Google Scholar 

  39. Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H, Alfonso ZC, Fraser JK, Benhaim P, Hedrick MH (2002) Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 13:4279–4295

  40. Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, Benhaim P, Lorenz HP, Hedrick MH (2001) Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 7:211–228

    Article  CAS  PubMed  Google Scholar 

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Acknowledgment

This work (NRF-2012R1A2A2A01012263) was supported by Mid-career Researcher Program through NRF grant funded by the MEST and a faculty research grant of Yonsei University College of Medicine (6-2008-0234).

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The authors declare no conflict of interests.

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Correspondence to Jin Woo Lee.

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Editor: T. Okamoto

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Jang, Y., Koh, Y.G., Choi, Y. et al. Characterization of adipose tissue-derived stromal vascular fraction for clinical application to cartilage regeneration. In Vitro Cell.Dev.Biol.-Animal 51, 142–150 (2015). https://doi.org/10.1007/s11626-014-9814-6

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Keywords

  • Bone marrow concentration
  • Adipose tissue-derived stromal vascular fraction
  • Chondrogenesis
  • Cartilage regeneration