Schiozer Wandir Antônio, Gemperli Rolf, Mühlbauer Wolfgang, Munhoz Alexandre Mendonça, Ferreira Marcus Castro
Acta Cir Bras. 2013 Dec;28(12):824-32. doi: 10.1590/s0102-86502013001200004.
To assess the viability of cultured epithelium and preserved by freezing for periods varying from one month to one year.
Samples of cultured epithelium were incubated in cryoprotectant medium (Group A), packed in aluminum envelopes and packed in polystyrene boxes. The boxes were subjected to a temperature of-70 ºC. After freezing for a period of time ranging from one to 12 months, cultured epithelial samples were assessed for their viability by vital staining (Trypan blue) and metabolic analysis based on glucose consumption and lactate production. Samples of not frozen cultured epithelium (Group B) were also tested for viability and the results obtained were used as comparison parameter for the variation of viability.
Statistical analysis between the group A and B indicate that the mean age of the donors (p=0.51) and the culture time (p=1.18) showed no statistical difference. In 30 days we obtained 37% of the original viability of cultured epithelium, 25% at six months and one year, less than 15%. This trend was confirmed statistically with a reduction of approximately 1.8% of the original viability epithelium cultured every 30 days of storage. In the analysis by lactate production, similar results were observed. In the analysis by the glucose consumption results were not significant. The viability indices show statistically significant difference between the group A and B (p<0.0001).
Although cryopreserved cultured epithelium showed significant reduction of viability, all samples remained viable. It was also found that the viability of cryopreserved cultured epithelial decreased as a function of storage time.
评估培养的上皮细胞经冷冻保存1个月至1年期间的活力。
将培养的上皮细胞样本置于冷冻保护培养基中孵育（A组），装入铝制封套，再放入聚苯乙烯盒中。将盒子置于-70ºC的温度下。冷冻1至12个月后，通过活细胞染色（台盼蓝）以及基于葡萄糖消耗和乳酸生成的代谢分析来评估培养的上皮细胞样本的活力。未冷冻的培养上皮细胞样本（B组）也进行活力测试，所得结果用作活力变化的比较参数。
A组和B组之间的统计分析表明，供体的平均年龄（p = 0.51）和培养时间（p = 1.18）无统计学差异。在30天时，培养的上皮细胞的活力为原始活力的37%，6个月和1年时为25%，1年时不到15%。每储存30天，培养的上皮细胞原始活力约降低1.8%，这一趋势经统计学证实。在乳酸生成分析中，观察到类似结果。在葡萄糖消耗分析中，结果不显著。活力指数显示A组和B组之间存在统计学显著差异（p < 0.0001）。
尽管冷冻保存的培养上皮细胞活力显著降低，但所有样本仍保持活力。还发现冷冻保存的培养上皮细胞的活力随储存时间而降低。
