Avaliação da viabilidade e fenótipo de células-tronco pulpares de dentes permanentes considerando as diferenças passagens celulares após criopreservação

Abstract

The aim of the study was to evaluate the cell viability, quantity, morphology and phenotype of pulp cells from permanent teeth (DPSCs) in different passages after three months of preservation at -80ºC. The DPSCs passed from sound third molars through the tissue explant technique and were expanded in culture, with two cryogenic tubes from passages 3, 6, 10 and 12 being frozen. After 3 months, the DPSCs were thawed and seeded in 96-well plates (20,000 cells/well) for viability assessment (MTT, n=9), 24 wells (50,000 cells/well) for morphological analysis, cell count through scanning electron microscopy (SEM, n=2) and in a petri dish 25 cm2 (100,000 cells/plate) for flow cytometric analysis of CD44, CD146, STRO-1 and CD45 herds. Data analysis used the ANOVA test, complemented by Tukey (p<0.05). The evaluation of viability and cell count showed higher values for #6 followed by #10. For the evaluation of the phenotype, it was observed that the #3 presented greater marking of CD146 and STRO-1 (means of 32.15% and 78.76% respectively), followed by #6 (means of 18.56% and 57 .96% respectively). Passages 10 and 12 presented minor markings (means of 4.66% and 5.38% for CD146 and 14.74% and 10.87% for STRO-1, respectively), with no statistical difference between them (p> 0.05). It was concluded that the #6 and #10 DPSCs had the best results regarding cell viability, quantity and morphology after 3 months of cryopreservation. In addition, a greater amount of undifferentiated cells were observed for #3 and #6.