Lindemann Daniele, Werle Stefanie B, Steffens Daniela, Garcia-Godoy Franklin, Pranke Patricia, Casagrande Luciano
Department of Pediatric Dentistry, School of Dentistry, Federal University of Rio Grande do Sul, Porto Alegre, Brazil.
School of Pharmacy, Federal University of Rio Grande do Sul, Porto Alegre, Brazil.
Arch Oral Biol. 2014 Sep;59(9):970-6. doi: 10.1016/j.archoralbio.2014.04.008. Epub 2014 May 2.
The aim of this study was to isolate and cultivate cells from the pulp of 7-day-cryopreserved intact deciduous human teeth and evaluate the effect of cryopreservation on dental pulp stem cell (DPSC) characteristics.
Twenty-six deciduous teeth were collected and allocated in two groups: immediate cell isolation (non-cryopreserved group) and intact cryopreserved (cryopreserved group). The teeth were cryopreserved in dimethylsulfoxide solution and recovered after 7 days. The success rate of isolation, proliferation, surface markers (CD14, CD29, CD34, CD45, CD73, CD90, and HLA-DR), differentiation capacity, and morphology were evaluated.
Isolation success rate was 61% and 30% for the non-cryopreserved and cryopreserved groups, respectively. There were no statistical differences between the groups for the tested surface markers. The cells in both groups were capable of differentiating into three mesenchymal lineages. No statistical differences between the groups were observed through the time course proliferation assay (0, 1, 3, 5, and 7 days); however, the mean time between isolation and the fifth passage was shorter for the non-cryopreserved group (p=0.035). The morphology of the cells was considered altered in the cryopreserved group.
DPSCs were obtained from cryopreserved intact deciduous teeth without changes in the immunophenotypical characteristics and differentiation ability; however, lower culture rates, proliferation potential, and morphological alterations were observed in relation to the control group.
本研究旨在从冷冻保存7天的完整人乳牙牙髓中分离并培养细胞,并评估冷冻保存对牙髓干细胞(DPSC)特性的影响。
收集26颗乳牙并分为两组:立即细胞分离(非冷冻保存组)和完整冷冻保存(冷冻保存组)。将牙齿置于二甲基亚砜溶液中冷冻保存,7天后复苏。评估分离成功率、增殖能力、表面标志物(CD14、CD29、CD34、CD45、CD73、CD90和HLA-DR)、分化能力及形态。
非冷冻保存组和冷冻保存组细胞分离成功率分别为61%和30%。两组间检测的表面标志物无统计学差异。两组细胞均能分化为三个间充质谱系。通过时间进程增殖试验(0、1、3、5和7天)未观察到两组间有统计学差异;然而,非冷冻保存组从分离到第五代的平均时间较短(p=0.035)。冷冻保存组细胞形态被认为发生了改变。
从冷冻保存的完整乳牙中获得了牙髓干细胞,其免疫表型特征和分化能力未发生改变;然而,与对照组相比,观察到较低的培养率、增殖潜能及形态改变。