CRYOPRESERVATION OF ISOLATED RAT HEPATOCYTES - A CRITICAL-EVALUATION OF FREEZING AND THAWING CONDITIONS
CRYOPRESERVATION OF ISOLATED RAT HEPATOCYTES - A CRITICAL-EVALUATION OF FREEZING AND THAWING CONDITIONS
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DOI：
10.1016/0011-2240(88)90040-5
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发表时间：
1988-08-01
期刊：
影响因子：
2.7
通讯作者：
GUILLOUZO, A
中科院分区：
文献类型：
--
作者：
CHESNE, C;GUILLOUZO, A
评估了成年大鼠肝细胞冷冻保存的各种参数，包括冷冻保护介质成分的性质和比例、冷却速率和解冻介质的组成。通过在含有 16% 二甲基亚砜的 L15 培养基中以 3°C 的速率冷冻，获得了能够在培养物中存活的最高百分比的细胞。 C/min，并向解冻培养基中添加 0.8 M 葡萄糖。超过 50% 的肝细胞能够在细胞分离后立即贴壁，在冷冻后保持这种特性并在原代培养物中存活。接种前可通过在 Percoll 层上离心来消除死细胞。在培养中，冷冻细胞表现出与未冷冻细胞相似的形态，24小时后，其蛋白质分泌率仅降低约40%。
Various parameters, including the nature and proportion of the constituents of the cryoprotective medium, the cooling rate, and the composition of the thawing medium, were evaluated for the cryopreservation of adult rat hepatocytes. The highest percentage of cells able to survive in culture was obtained by freezing in L15 medium containing 16% dimethyl sulfoxide, at a rate of 3.degree. C/min, and by adding 0.8 M glucose to the thawing medium. More than 50% of hepatocytes capable of attachment just after cell isolation kept this property after freezing and survived in primary culture. Dead cells could be eliminated before seeding by centrifugation on a Percoll layer. In culture, frozen cells exhibited a morphology similar to that of unfrozen cells and after 24 hr their protein secretion rate was reduced by only about 40%.