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首页> 外文期刊>Biopreservation and biobanking >Serum-Free Cryopreservation of Primary Rat Hepatocytes in a Modified Cold Storage Solution: Improvement of Cell Attachment and Function
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Serum-Free Cryopreservation of Primary Rat Hepatocytes in a Modified Cold Storage Solution: Improvement of Cell Attachment and Function

机译：修饰冷储存溶液中原代大鼠肝细胞的无血清冷冻保存：细胞附着和功能的改善

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Aims: The use of primary hepatocytes in pharmacological and toxicological research as well as for clinical and biotechnological applications requires adequate storage options for these cells. However, hepatocytes are very susceptible to cryopreservation injury. Based on experience in hypothermic storage of hepatocytes, we, in this study, aimed to optimize hepatocyte cryopreservation. Materials and Methods: Rat hepatocytes were cryopreserved in serum-containing cell culture medium or in serum-free solutions optimized for hypothermic storage, all with 10% dimethyl sulfoxide, using a standard protocol (-1 degrees C/min in a controlled-rate freezer). After rapid thawing, cells were seeded in supplemented Leibovitz-15 cell culture medium without further purification steps. Cell attachment and metabolic activity were assessed. Results: Cell attachment (37%15% vs. 9%+/- 7% of noncryopreserved control cells) and metabolic activity (resazurin reduction: 47%+/- 23% vs. 25%+/- 8%; glucose release: 44%+/- 6% vs. 15%+/- 7%; and urea production: 31%+/- 16% vs. 5%+/- 8%) were significantly higher after cryopreservation in the new solution compared to cryopreservation in cell culture medium. Experiments with modified solutions suggested that the protective effect of the new solution is multifactorial. Conclusions: In summary, significant improvement of cell attachment and function compared to cell culture medium was achieved after cryopreservation in serum-free hepatocyte cold storage solution.
机译：目的：在药理和毒理学研究中使用原发性肝细胞以及临床和生物技术应用需要对这些细胞进行足够的储存选择。然而，肝细胞非常易于冷冻保存损伤。在本研究中，基于肝细胞的低温储存经验，旨在优化肝细胞冷冻保存。材料和方法：在含血清细胞培养基中冷冻保存的大鼠肝细胞，用于低温储存的无血清溶液，所有配备10％二甲基磺氧化亚砜，使用标准方案（在受控速率冷冻机中为-1℃/ min ）。在快速解冻之后，将细胞接种在补充的Leibovitz-15细胞培养基中，无需进一步纯化步骤。评估细胞附着和代谢活性。结果：电池附着（37％15％vs.9％+ / - 7％的非髓等金属化对照细胞）和代谢活性（转塔唑素降低：47％+ / - 23％与25％+ / - 8％;葡萄糖释放： 44％+ / - 6％与15％+ / - 7％;和尿素产量：与冷冻保存相比，在新解决方案中冷冻保存后，31％+ / - 16％vs.5％+ / - 8％）显着提高在细胞培养基中。改性解决方案的实验表明新解决方案的保护作用是多因素。结论：总之，在无血清肝细胞冷储存溶液中的冷冻保存后实现了与细胞培养基相比细胞附着和功能的显着改善。

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