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Chondrogenic differentiation of human mesenchymal stem cells: a comparison between micromass and pellet culture systems

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Abstract

High-density cell culture is pivotal for the chondrogenic differentiation of human mesenchymal stem cells (hMSCs). Two high-density cell culture systems, micromass and pellet culture, have been used to induce chondrogenic differentiation of hMSCs. In micromass culture, the induced-cartilage tissues were larger, more homogenous and enriched in cartilage-specific collagen II but the fibrocartilage-like feature, collagen I, and hypertrophic chondrocyte feature, collagen X, were markedly decreased compared to those in pellet culture. Furthermore, real time RT-PCR analysis demonstrated that collagen II and aggrecan mRNA were up-regulated while collagen X and collagen I mRNA were down-regulated in micromass culture. Thus, the micromass culture system is a promising tool for in vitro chondrogenic studies.

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References

  • Djouad F, Mrugala D, Noel D et al (2006) Engineered mesenchymal stem cells for cartilage repair. Regen Med 1:529–537

    Article  CAS  PubMed  Google Scholar 

  • Erickson GR, Gimble JM, Franklin DM et al (2002) Chondrogenic potential of adipose tissue-derived stromal cells in vitro and in vivo. Biochem Biophys Res Commun 290:763–769

    Article  CAS  PubMed  Google Scholar 

  • Fehrer C, Brunauer R, Laschober G et al (2007) Reduced oxygen tension attenuates differentiation capacity of human mesenchymal stem cells and prolongs their lifespan. Aging Cell 6:745–757

    Article  CAS  PubMed  Google Scholar 

  • Johnstone B, Hering TM, Caplan AI et al (1998) In vitro chondrogenesis of bone marrow-derived mesenchymal progenitor cells. Exp Cell Res 238:265–272

    Article  CAS  PubMed  Google Scholar 

  • Kafienah W, Mistry S, Dickinson SC et al (2007) Three-dimensional cartilage tissue engineering using adult stem cells from osteoarthritis patients. Arthritis Rheum 56:177–187

    Article  PubMed  Google Scholar 

  • Kanichai M, Ferguson D, Prendergast PJ et al (2008) Hypoxia promotes chondrogenesis in rat mesenchymal stem cells: a role for AKT and hypoxia-inducible factor (HIF)-1alpha. J Cell Physiol 216:708–715

    Article  CAS  PubMed  Google Scholar 

  • Ke H, Wang P, Yu W et al (2009) Derivation, characterization and gene modification of cynomolgus monkey mesenchymal stem cells. Differentiation 77:256–262

    Article  CAS  PubMed  Google Scholar 

  • Malladi P, Xu Y, Chiou M et al (2006) Effect of reduced oxygen tension on chondrogenesis and osteogenesis in adipose-derived mesenchymal cells. Am J Physiol Cell Physiol 290:C1139–C1146

    Article  CAS  PubMed  Google Scholar 

  • Mello MA, Tuan RS (1999) High density micromass cultures of embryonic limb bud mesenchymal cells: an in vitro model of endochondral skeletal development. In Vitro Cell Dev Biol Anim 35:262–269

    Article  CAS  PubMed  Google Scholar 

  • Mueller MB, Tuan RS (2008) Functional characterization of hypertrophy in chondrogenesis of human mesenchymal stem cells. Arthritis Rheum 58:1377–1388

    Article  CAS  PubMed  Google Scholar 

  • Murdoch AD, Grady LM, Ablett MP et al (2007) Chondrogenic differentiation of human bone marrow stem cells in transwell cultures: generation of scaffold-free cartilage. Stem Cells 25:2786–2796

    Article  CAS  PubMed  Google Scholar 

  • Pelttari K, Steck E, Richter W (2008) The use of mesenchymal stem cells for chondrogenesis. Injury 39(Suppl 1):S58–S65

    Article  PubMed  Google Scholar 

  • Penick KJ, Solchaga LA, Welter JF (2005) High-throughput aggregate culture system to assess the chondrogenic potential of mesenchymal stem cells. Biotechniques 39:687–691

    Article  CAS  PubMed  Google Scholar 

  • Pittenger MF, Mackay AM, Beck SC et al (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147

    Article  CAS  PubMed  Google Scholar 

  • Scharstuhl A, Schewe B, Benz K et al (2007) Chondrogenic potential of human adult mesenchymal stem cells is independent of age or osteoarthritis etiology. Stem Cells 25:3244–3251

    Article  CAS  PubMed  Google Scholar 

  • Sundelacruz S, Kaplan DL (2009) Stem cell- and scaffold-based tissue engineering approaches to osteochondral regenerative medicine. Semin Cell Dev Biol 20:646–655

    Article  CAS  PubMed  Google Scholar 

  • Takagi M, Umetsu Y, Fujiwara M et al (2007) High inoculation cell density could accelerate the differentiation of human bone marrow mesenchymal stem cells to chondrocyte cells. J Biosci Bioeng 103:98–100

    Article  CAS  PubMed  Google Scholar 

  • Tare RS, Howard D, Pound JC et al (2005) Tissue engineering strategies for cartilage generation—micromass and three dimensional cultures using human chondrocytes and a continuous cell line. Biochem Biophys Res Commun 333:609–621

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (No. 30971587), the Science and Technology Planning Project of Guangdong Province (No. 2008B030301131) and the Yat-Sen Scholarship for Young Scientists.

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Correspondence to Dongsheng Huang.

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Zhang, L., Su, P., Xu, C. et al. Chondrogenic differentiation of human mesenchymal stem cells: a comparison between micromass and pellet culture systems. Biotechnol Lett 32, 1339–1346 (2010). https://doi.org/10.1007/s10529-010-0293-x

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  • DOI: https://doi.org/10.1007/s10529-010-0293-x

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