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Five percent oxygen tension is not beneficial for neocartilage formation in scaffold-free cell cultures

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Abstract

We have investigated whether 5% oxygen tension (O2) is beneficial for neocartilage formation when chondrocytes are cultured in transwell-COL inserts. Six million bovine primary chondrocytes were cultured in an insert with DMEM supplemented with 10% fetal bovine serum and antibiotics, with or without glucosamine sulphate (GS) in a 5% or 20% O2 environment for 2, 4, or 6 weeks. The samples were collected for the histological staining of proteoglycans (PGs) and type II collagen, quantitative reverse transcription with the polymerase chain reaction (RT-PCR) analyses of the mRNA expression of aggrecan and procollagen α1(II), procollagen α2(I) and hyaluronan synthase 2, quantitation of PGs, and agarose gel electrophoresis. Neocartilage produced at 20% O2 appeared larger than that at 5% O2. Histological staining showed that more PGs and type II collagen and better native cartilage structure were produced at 20% than at 5% O2. The thickness of neocartilage increased during the culture period. Quantitative RT-PCR showed that the procollagen α1(II) mRNA expression level was significantly higher at 20% than at 5% O2. However, no significant difference in gene expression and PG content was found between control and GS-treated cultures at either 20% or 5% O2. Thus, in contrast to monolayer cultures, engineered cartilage from scaffold-free cultured chondrocytes at 20% O2 produced better extracellular matrix (ECM) than that at 5% O2. PGs were mainly large. Exogenous GS was not beneficial for the ECM in scaffold-free chondrocyte cultures.

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Acknowledgements

We gratefully thank Elina Reinikainen, Eija Rahunen, and Kari Kotikumpu for their excellent assistance in the laboratory.

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Authors and Affiliations

  1. Department of Biosciences, Biocenter Kuopio, University of Eastern Finland, PO Box 1627, Yliopistoranta 1F, 70211, Kuopio, Finland

    Chengjuan Qu & Mikko J. Lammi

  2. Department of Biosciences, University of Eastern Finland, Kuopio, Finland

    Heli Lindeberg & Janne H. Ylärinne

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  1. Chengjuan Qu
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  2. Heli Lindeberg
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  3. Janne H. Ylärinne
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  4. Mikko J. Lammi
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Correspondence to Chengjuan Qu.

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This study was funded by the Paavo Nurmi Foundation and Academy of Finland (128177). This research was also supported by the strategic funding of the University of Eastern Finland.

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Qu, C., Lindeberg, H., Ylärinne, J.H. et al. Five percent oxygen tension is not beneficial for neocartilage formation in scaffold-free cell cultures. Cell Tissue Res 348, 109–117 (2012). https://doi.org/10.1007/s00441-012-1366-z

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  • DOI: https://doi.org/10.1007/s00441-012-1366-z

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