Wang Xin, Hua Tse-Chao, Sun Da-Wen, Liu Baolin, Yang Guanghui, Cao Yilin
Institute of Cryomedicine, Shanghai University of Science and Technology, Shanghai 200093, China.
Cryobiology. 2007 Aug;55(1):60-5. doi: 10.1016/j.cryobiol.2007.05.006. Epub 2007 Jun 9.
Cryopreservation of tissue-engineered human dermal replacement plays an important role in skin tissue engineering and skin banking. With the inspection of electronic scanning microscope and viability evaluation by Trypan Blue staining assay and the tetrazolium salt, MTT (3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay, this study investigated the toxicity of Me(2)SO to dermal fibroblasts and effects of cryoprotectant concentration and cooling rate on the viability of dermal replacement. The results demonstrated that the Me(2)SO toxicity to fibroblasts was affected by the exposure time, temperature, and concentration. Furthermore adding cryoprotectant solution at low temperature of 4 degrees C significantly reduced the toxic effect on the tissue-engineered dermal equivalent. An optimal cryopreservation protocol consisting of cooling rate at 1 degrees Cmin(-1) in 10% (V/V) Me(2)SO was derived, with the viability of studied dermal equivalent treated by this protocol being 75% of that of fresh control. The micrograph obtained by electronic scanning microscope also confirmed this result.
组织工程化人真皮替代物的冷冻保存在皮肤组织工程和皮肤库中发挥着重要作用。通过电子扫描显微镜检查以及台盼蓝染色法和四氮唑盐MTT（3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐）法进行活力评估，本研究调查了二甲基亚砜（Me(2)SO）对真皮成纤维细胞的毒性以及冷冻保护剂浓度和降温速率对真皮替代物活力的影响。结果表明，Me(2)SO对成纤维细胞的毒性受暴露时间、温度和浓度的影响。此外，在4℃低温下添加冷冻保护剂溶液可显著降低对组织工程化真皮替代物的毒性作用。得出了一种最佳冷冻保存方案，即在10%（V/V）Me(2)SO中以1℃/min(-1)的降温速率进行冷冻，经该方案处理的研究用真皮替代物的活力为新鲜对照的75%。电子扫描显微镜获得的显微照片也证实了这一结果。
