Studies of chemical toxicity to fresh and cryopreserved rat hepatocytes.
Studies of chemical toxicity to fresh and cryopreserved rat hepatocytes.
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对新鲜和冷冻保存的大鼠肝细胞的化学毒性研究。
DOI：
10.1016/0041-008x(89)90341-4
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发表时间：
1989
影响因子：
3.8
通讯作者：
Powis,G
中科院分区：
文献类型：
--
作者：
Santone,KS;Melder,DC;Powis,G
分离的肝细胞可用于研究外源性化学物质的代谢和肝毒性机制。使用人肝细胞的一个问题是正常人肝脏的有限和不规则的可用性。冷冻保存可以提供一种有用的方法来储存肝细胞，直到它们被需要。作为使用人肝细胞的初步步骤，我们比较了新鲜大鼠肝细胞和如前所述冷冻保存的大鼠肝细胞的原代培养物对化学毒物的毒性反应（G. Powis，K. S. Santone，D. C.梅尔德湖托马斯，D.摩尔和T. J. Wilke，1987年。药物代谢Dispos.15，826）。培养24小时后，冻存肝细胞的平板接种效率为非冻存肝细胞的75%。与培养的非冻存肝细胞相比，培养的冻存肝细胞自发乳酸脱氢酶释放略有增加。类似的毒性化学物质诱导的乳酸脱氢酶释放的增加发生在培养的冷冻保存的noncryopreserved肝细胞。乳酸脱氢酶释放的50%有效浓度（EC 50）（±SE，n = 3份制备物），氯丙嗪、氯化镉和甲萘醌（2-甲基-1，4-萘醌）分别为235 ± 20和215 ± 30 μm、200 ± 5和272 ± 23 μm和24 ± 7和44 ± 8 μm。在冻存和非冻存培养的肝细胞中，氯丙嗪、氯化镉和甲萘醌的细胞内谷胱甘肽耗竭的EC 50值分别为200 ± 8和235 ± 8 μm、242 ± 19和213 ± 7 μm以及22 ± 2和21 ± 3 μm。结果表明，冷冻保存为化学毒性研究提供了一种实用的大鼠肝细胞保存方法。
Isolated hepatocytes are useful for studying the metabolism and mechanisms of hepatic toxicity of foreign chemicals. A problem with using human hepatocytes is the limited and irregular availability of normal human liver. Cryopreservation could provide a useful way of storing hepatocytes until they are needed. As a preliminary step to using human hepatocytes we have compared the toxic response to chemical toxicants of primary cultures of fresh rat hepatocytes and rat hepatocytes cryopreserved as previously described (G. Powis, K. S. Santone, D. C. Melder, L. Thomas, D. J. Moore, and T. J. Wilke, 1987. Drug Metab. Dispos.15, 826). After 24 hr in culture the cryopreserved hepatocytes had a plating efficiency 75% that of noncryopreserved hepatocytes. The cultured cryopreserved hepatocytes showed a small increase in spontaneous lactate dehydrogenase release compared to that of cultured noncryopreserved hepatocytes. A similar toxic chemical-induced increase in lactate dehydrogenase release occurred in the cultured cryopreserved as in the noncryopreserved hepatocytes. The 50% effective concentrations (EC50) for lactate dehydrogenase release (±SE, n = 3 preparations) from cultured cryopreserved and noncryopreserved hepatocytes for chlorpromazine were 235 ± 20 and 215 ± 30 μm, for cadmium chloride 200 ± 5 and 272 ± 23 μm, and for menadione (2-methyl-1,4-naphthoquinone) 24 ± 7 and 44 ± 8 μm, respectively. The EC50 values for intracellular glutathione depletion in cultured cryopreserved and noncryopreserved hepatocytes were for chlopromazine 200 ± 8 and 235 ± 8 μm, for cadmium chloride 242 ± 19 and 213 ± 7 μm, and for menadione 22 ± 2 and 21 ± 3 μm, respectively. The results show that cryopreservation offers a practical way of storing rat hepatocytes for studies of chemical toxicity.