Summary
We have investigated the viability and function of cells in cartilage slices after various methods of preservation, and have examined the viability of cells by measuring the incorporation of Na2 35SO4 at different concentrations, temperatures and times of exposure to cryopreservatives. We have assessed the viability of cells when exposed to prefreezing temperatures, and after preservation at −80° C. The best survival rate was found to be with a concentration of cryopreservatives of approximately 10%, with pre-freeze exposure for four hours at 4° C. In the stage cooling technique, the best initial cooling was at −30° C for 30 minutes, followed by rapid cooling of the cartilage to −80° C. The best survival rate for cryopreserved cartilage in 10% DMSO was, on average, 19% in intact slices and 34% when holes had been made in the slices. Proteoglycan synthesis after thawing appeared normal, and proteoglycan labelled after 48 hours in culture also showed a normal pattern.
Résumé
Nous avons étudié la viabilité et la fonction cellulaire de coupes de cartilage après diverses méthodes de conservation et nous avons évalué la viabilité des cellules en mesurant l'absorption du Na2 35SO4 à différentes concentrations, températures et durées d'exposition aux cryo-préservateurs. Nous avons évalué la viabilité des cellules exposées aux températures de pré-congélation et après conservation à −80° C. Nous avons trouvé que le meilleur pourcentage de survie était obtenu avec une concentration de cryo-préservateurs d'environ 10%, avec une pré-congélation de 4 heures à 4° C. Dans la technique de refroidissement par étapes, le meilleur refroidissement initial est de −30° C pendant 30 minutes, suivi d'un refroidissement rapide du cartilage jusqu'a −80° C. Le meilleur taux de survie pour le cartilage cryo-préservé dans 10% de DMSO a été, en moyenne, de 19% dans les coupes intactes et de 34% quand des perforations ont été pratiquées dans les coupes. La synthèse du protéoglycan après décongélation paraît normale et le protéoglycan marqué, après une culture de 48 heures, s'avère également normal.
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Kawabe, N., Yoshinao, M. Cryopreservation of cartilage. International Orthopaedics 14, 231–235 (1990). https://doi.org/10.1007/BF00178751
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DOI: https://doi.org/10.1007/BF00178751