冷冻猪肝细胞在生物型人工肝系统中的生存状态研究

Living states of cryopreserved porcine hepatocytes in biological artificial liver support system

目的 研究冷冻复温后猪肝细胞在生物型人工肝支持系统(BALSS)中的生存状态。方法 用酶法从中国实验用小型猪肝脏分离出猪肝细胞;将1×10 1 0 猪肝细胞冷冻保存在-196℃液氮中;1个月后复温,将等量的冷冻和新鲜猪肝细胞分别培养在BALSS中,比较其形态、存活率和白蛋白、尿素、葡萄糖合成功能及对利多卡因的转化功能。结果 冷冻保存的猪肝细胞,复温后其存活率较高;在培养于BALSS时其生化功能、利多卡因转化功能及超微结构与新鲜肝细胞类似。结论 用该冻存方法保存的猪肝细胞可试用于BALSS系统。

Objective To study the living states of cryopreserved porcine hepatocytes under the support of biological artificial liver system. Methods Porcine hepatocytes were isolated from Chinese experimental miniature swines by collagenase digestion and stored with aliquots of 1×1 010 cells at -196 ℃ liquid nitrogen. After 1 month, the stored cells were resuscitated and cultured in bioartificial liver support system (BALSS). Fresh hepatocytes at same amount were served as control. The morphology, survival rate, and functions of synthesizing albumin, urea and glucose and transforming lidocaine were observed. Results The survival rate of the frozen hepatocytes was high. There was no obvious difference of the two group in the cell function and microstructure. Conclusion The results indicate that cryopreserved hepatocytes can be used in BALSS.