Hypertonic conditions enhance cartilage formation in scaffold-free primary chondrocyte cultures
2014 (English)In: Cell and Tissue Research, ISSN 0302-766X, E-ISSN 1432-0878, ISSN 1432-0878, Vol. 358, no 2, 541-550 p.Article in journal (Refereed) Published
The potential of hypertonic conditions at in vivo levels to promote cartilage extracellular matrix accumulation in scaffold-free primary chondrocyte cultures was investigated. Six million bovine primary chondrocytes were cultured in transwell inserts in low glucose (LG), high glucose (HG), or hypertonic high glucose (HHG) DMEM supplemented with fetal bovine serum, antibiotics, and ascorbate under 5 % or 20 % O2 tension with and without transforming growth factor (TGF)-β3 for 6 weeks. Samples were collected for histological staining of proteoglycans (PGs) and type II collagen, analysis by quantitative reverse transcription plus the polymerase chain reaction (RT-PCR) of mRNA expression of aggrecan and procollagen α1 (II) and of Sox9 and procollagen α2 (I), and quantitation of PGs and PG separation in agarose gels. Cartilage tissues produced at 20 % O2 tension were larger than those formed at 5 % O2 tension. Compared with LG, the tissues grew to larger sizes in HG or HHG medium. Histological staining showed the strongest PG and type II collagen staining in cartilage generated in HG or HHG medium at 20 % O2 tension. Quantitative RT-PCR results indicated significantly higher expression of procollagen α1 (II) mRNA in cartilage generated in HHG medium at 20 % O2 tension compared with that in the other samples. TGF-β3 supplements in the culture medium provided no advantage for cartilage formation. Thus, HHG medium used at 20 % O2 tension is the most beneficial combination of the tested culture conditions for scaffold-free cartilage production in vitro and should improve cell culture for research into cartilage repair or tissue engineering.
Place, publisher, year, edition, pages
Berlin: Springer Berlin/Heidelberg, 2014. Vol. 358, no 2, 541-550 p.
cartilage, scaffold-free culture, oxygen tension, TGF-beta, bovine, tissue engineering
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Research subject cellforskning
IdentifiersURN: urn:nbn:se:umu:diva-103842DOI: 10.1007/s00441-014-1970-1ISI: 000344348500022PubMedID: 25107609OAI: oai:DiVA.org:umu-103842DiVA: diva2:815868