Vasconcelos Rodrigo Gadelha, Ribeiro Rodrigo Alves, Vasconcelos Marcelo Gadelha, Lima Kenio Costa, Barboza Carlos Augusto Galvão
Postgraduate Program in Oral Pathology, Federal University of Rio Grande do Norte, Natal, Rio Grande do Norte, Brazil.
Cell Tissue Bank. 2012 Aug;13(3):461-9. doi: 10.1007/s10561-011-9271-3. Epub 2011 Jul 21.
Cryopreservation aims to cease all biological functions of living tissues in a reversible and controlled manner, i.e., to permit the recovery of cells by maintaining a high degree of their viability and functional integrity. The objective of this study was to evaluate in vitro the influence of cryopreservation on undifferentiated mesenchymal cells derived from the periodontal ligament of human third molars. Mesenchymal cells were isolated from six healthy teeth and cultured in α-MEM medium supplemented with antibiotics and 15% FBS in a humid atmosphere with 5% CO(2) at 37°C. The cells isolated from each tooth were divided into two groups: group I (fresh, non-cryopreserved cells) was immediately cultured, and group II was submitted to cryopreservation for 30 days. The rates of cell adhesion and proliferation were analyzed in the two groups by counting the cells adhered to the wells at 24, 48 and 72 h after plating. The number of cells per well was obtained by counting viable cells in a hemocytometer using the Trypan blue exclusion method. Differences between groups at each time point were evaluated by the Wilcoxon test. The Friedman test was used to determine differences between time points and, if detected, the Wilcoxon test with Bonferroni correction was applied. The results showed no significant difference in the in vitro growth capacity of undifferentiated mesenchymal cells between the two groups. In conclusion, cryopreservation for 30 days had no influence on periodontal ligament mesenchymal cells.
冷冻保存旨在以可逆且可控的方式停止活组织的所有生物功能，即通过维持细胞的高度活力和功能完整性来使其得以复苏。本研究的目的是在体外评估冷冻保存对源自人类第三磨牙牙周韧带的未分化间充质细胞的影响。从六颗健康牙齿中分离出间充质细胞，并在补充有抗生素和15%胎牛血清的α-MEM培养基中，于37°C、5%二氧化碳的潮湿气氛中培养。从每颗牙齿分离出的细胞分为两组：第一组（新鲜的、未冷冻保存的细胞）立即进行培养，第二组进行30天的冷冻保存。通过在接种后24、48和72小时计数贴壁于孔中的细胞，分析两组细胞的黏附率和增殖率。使用台盼蓝排斥法通过血细胞计数板计数活细胞来获得每孔中的细胞数量。通过Wilcoxon检验评估各时间点两组之间的差异。使用Friedman检验确定时间点之间的差异，若检测到差异，则应用经Bonferroni校正的Wilcoxon检验。结果显示两组未分化间充质细胞的体外生长能力无显著差异。总之，30天的冷冻保存对牙周韧带间充质细胞没有影响。
