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ROCK Y-27632 Inhibiteur, L'acide Ascorbique Et Le Tréhalose Augmentent La Survie Des Cellules Souches Mésenchymateuses De La Gelée De Wharton Humaine Après Cryoconservation.

OBJECTIFS: Les cellules souches mésenchymateuses de la gelée Wharton sont de bons candidats pour une application dans différents aspects de la médecine régénérative, et leur stockage à long terme est important. Dans cette étude, les effets du tréhalose, de l'acide ascorbique et du Y-27632 sur la prolifération et le taux de survie de ces cellules après cryoconservation ont été étudiés. MATÉRIAUX ET MÉTHODES: Des cellules souches mésenchymateuses ont été isolées de la gelée de Wharton de cordon ombilical humain et congelées à l'aide d'un processus de refroidissement à vitesse lente. Différentes concentrations de tréhalose (35, 75 et 125 mM), d'acide ascorbique (0,06, 0,125, 0,25 et 0,5 mM) et de Y-27632 (10 μM) ont été utilisées pour traiter le milieu de culture et / ou compléter le milieu de congélation. L'évaluation de la viabilité cellulaire après décongélation a été réalisée en utilisant une coloration au bleu Trypan, et un dosage au MTT a été effectué pour mesurer le taux de prolifération cellulaire. RÉSULTATS: Nous avons observé une augmentation significative de la viabilité post-dauphin, une prolifération cellulaire accrue et une réduction du temps de doublement des cellules lorsque du tréhalose 75 mM, de l'acide ascorbique 0,25 et 0,5 mM et du Y-27632 10 mM ont été utilisés. De plus, une viabilité, une prolifération et une fixation accrues ont été observées après 24 heures de prétraitement avec ces agents cryoprotecteurs et lorsqu'ils ont été ajoutés à un milieu de congélation classique. CONCLUSIONS: L'utilisation de différents agents cryoprotecteurs dans les milieux de culture et de congélation pourrait être utile pour le stockage à long terme des cellules souches mésenchymateuses de la gelée Wharton.


Source: https://www.ncbi.nlm.nih.gov/pubmed/29957164


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