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In vitro cartilage formation of composites of synovium-derived mesenchymal stem cells with collagen gel

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Abstract

Graft implantation is one of the more popular procedures for repairing cartilage defects; however, sacrifices of the donor site have been an issue. Mesenchymal stem cells (MSCs) are a fascinating source for regenerative medicine because they can be harvested in a less invasive manner and are easily isolated and expanded, with multipotentiality including chondrogenesis. MSCs can be isolated from various adult mesenchymal tissues including synovium. Here, we attempted to form cartilage from the composites of synovium-derived MSCs with collagen gel in vitro. After 21 days of culture, the composites had increased their cartilage matrix, as demonstrated by toluidine blue staining and immunohistochemistry for type II collagen. The composites consisting of 5×107 and 108 cells/ml in gel were richer in proteoglycans than those consisting of lower cell densities. After 1 day, MSCs/gel composites contracted and the diameter decreased by 30%; however, they were stable thereafter. Round cells with short processes producing collagen fibrils showing a similar morphology to that of chondrocytes were seen in the composites by transmission electron microscopy. During composite culture, chondroitin sulfate and mRNA expression for cartilage-related genes increased, demonstrating cartilage maturation. Using an optimized method, we obtained cartilage discs with a diameter of 7 mm and a thickness of 500 μm. Our procedure should thus make it possible to produce a large cartilage matrix in vitro. The tissue engineering of autologous cartilage from the composites of synovium-derived MSCs with collagen gel in vitro for transplantation may be a future alternative to graft implantation for patients with cartilage defects.

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References

  • Barry F, Boynton R, Murphy M, Haynesworth S, Zaia J (2001) The SH-3 and SH-4 antibodies recognize distinct epitopes on CD73 from human mesenchymal stem cells. Biochem Biophys Res Commun 289:519–524

    Article  PubMed  CAS  Google Scholar 

  • Bell E, Ivarsson B, Merrill C (1979) Production of a tissue-like structure by contraction of collagen lattices by human fibroblasts of different proliferative potential in vitro. Proc Natl Acad Sci U S A 76:1274–1278

    Article  PubMed  CAS  Google Scholar 

  • Brent B (1992) Auricular repair with autogenous rib cartilage grafts: two decades of experience with 600 cases. Plast Reconstr Surg 90:355–374

    Article  PubMed  CAS  Google Scholar 

  • 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  PubMed  CAS  Google Scholar 

  • Colter DC, Sekiya I, Prockop DJ (2001) Identification of a subpopulation of rapidly self-renewing and multipotential adult stem cells in colonies of human marrow stromal cells. Proc Natl Acad Sci U S A 98:7841–7845

    Article  PubMed  CAS  Google Scholar 

  • De Bari C, Dell'Accio F, Tylzanowski P, Luyten FP (2001) Multipotent mesenchymal stem cells from adult human synovial membrane. Arthritis Rheum 44:1928–1942

    Article  PubMed  Google Scholar 

  • Ichinose S, Muneta T, Sekiya I, Itoh S, Aoki H, Tagami M (2003) The study of metal ion release and cytotoxicity in Co-Cr-Mo and Ti-Al-V alloy in total knee prosthesis - scanning electron microscopic observation. J Mater Sci Mater Med 14:79–86

    Article  PubMed  CAS  Google Scholar 

  • Liu H, Lee YW, Dean MF (1998) Re-expression of differentiated proteoglycan phenotype by dedifferentiated human chondrocytes during culture in alginate beads. Biochim Biophys Acta 1425:505–515

    PubMed  CAS  Google Scholar 

  • Maeda S, Miyabayashi T, Yamamoto JK, Roberts GD, Lepine AJ, Clemmons RM (2003) Temporal dynamic changes in synthesis of chondroitin sulfate isomers in canine articular chondrocyte culture. J Vet Med Sci 65:1373–1376

    Article  PubMed  CAS  Google Scholar 

  • Maurice H, Crone M, Watt I (1988) Synovial chondromatosis. J Bone Joint Surg Br 70:807–811

    PubMed  CAS  Google Scholar 

  • Mourao PA (1988) Distribution of chondroitin 4-sulfate and chondroitin 6-sulfate in human articular and growth cartilage. Arthritis Rheum 31:1028–1033

    Article  PubMed  CAS  Google Scholar 

  • Nishimura K, Solchaga LA, Caplan AI, Yoo JU, Goldberg VM, Johnstone B (1999) Chondroprogenitor cells of synovial tissue. Arthritis Rheum 42:2631–2637

    Article  PubMed  CAS  Google Scholar 

  • Ochi M, Uchio Y, Tobita M, Kuriwaka M (2001) Current concepts in tissue engineering technique for repair of cartilage defect. Artif Organs 25:172–179

    Article  PubMed  CAS  Google Scholar 

  • 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  PubMed  CAS  Google Scholar 

  • Prockop DJ (1997) Marrow stromal cells as stem cells for nonhematopoietic tissues. Science 276:71–74

    Article  PubMed  CAS  Google Scholar 

  • Sakaguchi Y, Sekiya I, Yagishita K, Ichinose S, Shinomiya K, Muneta T (2004) Suspended cells from trabecular bone by collagenase digestion become virtually identical to mesenchymal stem cells obtained from marrow aspirates. Blood 104:2728–2735

    Article  PubMed  CAS  Google Scholar 

  • Sekiya I, Colter DC, Prockop DJ (2001) BMP-6 enhances chondrogenesis in a subpopulation of human marrow stromal cells. Biochem Biophys Res Commun 284:411–418

    Article  PubMed  CAS  Google Scholar 

  • Sekiya I, Larson BL, Smith JR, Pochampally R, Cui JG, Prockop DJ (2002a) Expansion of human adult stem cells from bone marrow stroma: conditions that maximize the yields of early progenitors and evaluate their quality. Stem Cells 20:530–541

    Article  PubMed  Google Scholar 

  • Sekiya I, Vuoristo JT, Larson BL, Prockop DJ (2002b) In vitro cartilage formation by human adult stem cells from bone marrow stroma defines the sequence of cellular and molecular events during chondrogenesis. Proc Natl Acad Sci U S A 99:4397–4402

    Article  PubMed  CAS  Google Scholar 

  • Sekiya I, Larson BL, Vuoristo JT, Reger RL, Prockop DJ (2005) Comparison of effect of BMP−2, −4, and −6 on in vitro cartilage formation of human adult stem cells from bone marrow stroma. Cell Tissue Res 320:269–276

    Article  PubMed  CAS  Google Scholar 

  • Shinmei M, Miyauchi S, Machida A, Miyazaki K (1992) Quantitation of chondroitin 4-sulfate and chondroitin 6-sulfate in pathologic joint fluid. Arthritis Rheum 35:1304–1308

    PubMed  CAS  Google Scholar 

  • Tanzer RC (1959) Total reconstruction of the external ear. Plast Reconstr Surg 23:1–15

    Article  PubMed  CAS  Google Scholar 

  • Tsutsumi K, Shimakawa H, Kitagawa H, Sugahara K (1998) Functional expression and genomic structure of human chondroitin 6-sulfotransferase. FEBS Lett 441:235–241

    Article  PubMed  CAS  Google Scholar 

  • Uchimura K, Kadomatsu K, Nishimura H, Muramatsu H, Nakamura E, Kurosawa N, Habuchi O, El-Fasakhany FM, Yoshikai Y, Muramatsu T (2002) Functional analysis of the chondroitin 6-sulfotransferase gene in relation to lymphocyte subpopulations, brain development, and oversulfated chondroitin sulfates. J Biol Chem 277:1443–1450

    Article  PubMed  CAS  Google Scholar 

  • Yamauchi S, Mita S, Matsubara T, Fukuta M, Habuchi H, Kimata K, Habuchi O (2000) Molecular cloning and expression of chondroitin 4-sulfotransferase. J Biol Chem 275:8975–8981

    Article  PubMed  CAS  Google Scholar 

  • Yoshida K, Miyauchi S, Kikuchi H, Tawada A, Tokuyasu K (1989) Analysis of unsaturated disaccharides from glycosaminoglycuronan by high-performance liquid chromatography. Anal Biochem 177:327–332

    Article  PubMed  CAS  Google Scholar 

  • Zannettino AC, Harrison K, Joyner CJ, Triffitt JT, Simmons PJ (2003) Molecular cloning of the cell surface antigen identified by the osteoprogenitor-specific monoclonal antibody, HOP-26. J Cell Biochem 89:56–66

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

We thank Kenichi Shinomiya for supporting our studies, Yusuke Sakaguchi for help with the characterization of MSCs, Hideya Yoshimura for help with real-time PCR, Izumi Nakagawa for excellent technical assistance, Miyoko Ojima for expert help with histology, and Kelly Johanson for proofreading. Recombinant human BMP2 was from the Yamanouchi Pharmaceutical Company.

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Correspondence to Ichiro Sekiya.

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This study is supported in part by grants from the Japanese Society for the Promotion of Science (16591478), the Japanese Orthopaedics and Traumatology Foundation, and the Nakatomi Foundation to I.S., and the Japanese Society for the Promotion of Science (16591477), the Japanese Sports Medicine Foundation, the Japanese Latest Osteoarthritis Society, and the Center of Excellence Program for Frontier Research on Molecular Destruction and Reconstruction of Tooth and Bone in Tokyo Medical and Dental University to T.M.

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Yokoyama, A., Sekiya, I., Miyazaki, K. et al. In vitro cartilage formation of composites of synovium-derived mesenchymal stem cells with collagen gel. Cell Tissue Res 322, 289–298 (2005). https://doi.org/10.1007/s00441-005-0010-6

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  • DOI: https://doi.org/10.1007/s00441-005-0010-6

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