Determination of Viability of Human Cartilage Allografts by a Rapid and Quantitative Method Not Requiring Cartilage Digestion
Abstract:Fresh osteochondral allograft transplantation is increasingly used for the treatment of cartilage pathologies of the knee. It is believed that transplantation success depends on the presence of viable chondrocytes in the graft, but methods to evaluate graft viability require the isolation of chondrocytes by enzymatic digestion of the cartilage and/or the use of radioactive precursors. We have adapted the well-known cell viability assay based on the reduction of tetrazolium derivatives to evaluate cartilage viability. We took advantage from the histological properties of cartilage tissue and the fact that some tetrazolium derivatives (e.g., WST-1, XTT) give soluble reduction products that can permeate the hyaline cartilage matrix. We have validated this assay in human cartilage explants from arthrotomy interventions and deceased donors, measuring the reduced product in the explant supernatant. Using this method we have compared the performance of several culture media in cartilage viability. From those tested, DMEM supplemented with fetal bovine serum results in higher viability of the cartilage and the explants remain viable at least 15 days in culture at 37°C. Cartilage cells continued expressing chondrocyte-specific genes, suggesting the maintenance of chondrogenic phenotype. The described method offers a quantitative and convenient method to measure the viability of human cartilage grafts.
Document Type: Research Article
Publication date: July 1, 2008
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- Cell Transplantation publishes original, peer-reviewed research and review articles on the subject of cell transplantation and its application to human diseases. To ensure high-quality contributions from all areas of transplantation, separate section editors and editorial boards have been established. Articles deal with a wide range of topics including physiological, medical, preclinical, tissue engineering, and device-oriented aspects of transplantation of nervous system, endocrine, growth factor-secreting, bone marrow, epithelial, endothelial, and genetically engineered cells, among others. Basic clinical studies and immunological research papers are also featured. To provide complete coverage of this revolutionary field, Cell Transplantation will report on relevant technological advances, and ethical and regulatory considerations of cell transplants. Cell Transplantation is now an Open Access journal starting with volume 18 in 2009, and therefore there will be an inexpensive publication charge, which is dependent on the number of pages, in addition to the charge for color figures. This will allow work to be disseminated to a wider audience and also entitle the corresponding author to a free PDF, as well as prepublication of an unedited version of the manuscript.