Cryopreservation of isolated rat hepatocytes.
Cryopreservation of isolated rat hepatocytes.
复制标题
分离的大鼠肝细胞的冷冻保存。
DOI：
10.1007/bf02796340
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发表时间：
1982
期刊：
影响因子：
--
通讯作者：
Michalopoulos,G
中科院分区：
文献类型：
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作者：
Novicki,DL;Irons,GP;Strom,SC;Jirtle,R;Michalopoulos,G
将成年大鼠分离的实质肝细胞冷冻在含10%甘油、10%二甲基亚砜（DMSO）或20% DMSO的培养基中。在非冷冻细胞和在上述每种冷冻保护剂溶液中冷冻的细胞中比较了三种微粒体相关功能。冷冻在DMSO中保持细胞色素P-450和b5和NADPH-细胞色素C还原酶的水平更接近控制值比冷冻在甘油中。细胞冷冻，随后解冻和培养24小时失去了更大量的细胞色素P-450比非冷冻培养的细胞。细胞色素b5和还原酶的水平在冻融细胞保持接近控制值。使用DMSO作为冷冻保护剂可更好地维持冷冻后的细胞活力（台盼蓝染料排除和附着细胞的百分比）。二甲亚砜可保护肝细胞免受冷冻诱导的损伤，使许多活细胞附着在胶原包被的培养皿上，存活至少24 h，并仍保持对药物和致癌物代谢重要的酶的显著水平。
Isolated parenchymal hepatocytes from adult rats were frozen in media containing 10% glycerol, 10% dimethylsulfoxide (DMSO), or 20% DMSO. Three microsome-associated functions were compared in nonfrozen cells and cells frozen in each of the above cryoprotectant solutions. Freezing in DMSO maintains cytochromes P-450 and b5and NADPH-cytochrome C reductase at levels nearer to control values than does freezing in glycerol. Cells frozen and subsequently thawed and cultured for 24 h lose a greater amount of cytochrome P-450 than do nonfrozen cultured cells. The levels of cytochrome b5and reductase in frozen-thawed cells remain close to control values. Cell viability (trypan blue dye exclusion and percentage of attached cells) after freezing is maintained better using DMSO as a cryoprotectant. Dimethylsulfoxide protects the hepatocytes from freeze-induced damage to the extent that many viable cells attach to collagen-coated petri dishes, survive for at least 24 h, and still maintain significant levels of enzymes of importance to drug and carcinogen metabolism.