Viability and drug metabolism capacity of alginate-entrapped hepatocytes after cryopreservation
Viability and drug metabolism capacity of alginate-entrapped hepatocytes after cryopreservation
复制标题
海藻酸盐包埋肝细胞冻存后的活力和药物代谢能力
DOI：
10.1023/a:1007690009927
复制
发表时间：
2004
影响因子：
6.1
通讯作者：
A. Guillouzo
中科院分区：
文献类型：
--
作者：
Laure Rialland;C. Guyomard;M. Scotté;C. Chesné;A. Guillouzo
在目前的研究中，我们评估了不同物种的冷冻保存的肝细胞的活力和解毒酶能力，包括人，固定在海藻酸钙凝胶中。利用特定的底物，分别测定了异种代谢主要细胞色素P450(CYP)的乙氧基间苯二酚O-脱乙基酶、非那西丁脱乙基酶、戊氧基间苯二酚O-脱烷基酶、甲苯磺胺羟基酶、S-甲苯妥因羟基酶、右美沙芬脱甲基酶、硝苯地平氧化酶以及谷胱甘肽S转移酶的活性。冷冻保存后，固定化肝细胞的活性仅略有下降，大部分CYP相关的单加氧酶活性被很好地保存下来，表达水平接近未冷冻的肝细胞单层。相比之下，总GST活性下降了约50%。然而，就像CYP1A和3A相关的酶一样，大鼠GST仍然能够对典型的诱导剂做出反应。在未冷冻和冷冻的固定化肝细胞以及未冷冻的肝细胞单层中，倍数增加是相似的。储存期，即使超过一年，也不影响活性和功能。综上所述，冷冻保存后，藻酸盐包裹的肝细胞仍然具有很高的活性，并且能够表达大多数解毒酶，其表达水平接近于相应的未冷冻肝细胞单层中的表达水平，并且能够对典型的诱导剂做出反应。
In the present study we evaluated viability and detoxifying enzyme capacity of cryopreserved hepatocytes from various species, including man, immobilized in calcium alginate gels. Ethoxyresorufin O-deethylase, phenacetin deethylase, pentoxyresorufin O-dealkylase, tolbutamide hydroxylase, S-mephenytoin hydroxylase, dextromethorphan demethylase, and nifedipine oxidation corresponding to the major cytochromes P450 (CYP) involved in xenobiotic metabolism as well as whole glutathione S-transferase (GST) activity were measured using specific substrates and after exposure or not to prototypical inducers. After deep-freeze storage, viability of immobilized hepatocytes was only slightly reduced and most CYP-related monooxygenase activities were well preserved, being expressed at levels close to those measured in unfrozen hepatocyte monolayers. By contrast, total GST activity was decreased by around 50%. However, as did CYP1A- and 3A-related enzymes, rat GST remained capable of responding to prototypical inducers. The fold increases were comparable in unfrozen and frozen immobilized hepatocytes and in unfrozen hepatocyte monolayers. The duration of storage, even when exceeding one year, did not affect viability and functions. In conclusion, after cryopreservation, alginate-entrapped hepatocytes remain highly viable and capable of expressing most detoxifying enzymes at levels close to those expressed in corresponding unfrozen hepatocyte monolayers and of responding to prototypical inducers.
冷冻保存大鼠和狗肝细胞，用于研究外源代谢和激活。
DOI：
--
发表时间：
1987
期刊：
Drug metabolism and disposition: the biological fate of chemicals
影响因子：
--
作者：
Powis,G;Santone,KS;Melder,DC;Thomas,L;Moore,DJ;Wilke,TJ
通讯作者：
Wilke,TJ
有效冷冻保存和长期储存原代人肝细胞，恢复活力、分化和复制潜力。
影响因子：
3.3
作者：
Adams,RM;Wang,M;Crane,AM;Brown,B;Darlington,GJ;Ledley,FD
通讯作者：
Ledley,FD