Cell cryopreservation stops the biological activity of cells by placing them in the frozen state, and can be used to preserve cells without subculturing, which can cause contamination and genetic drift. However, the freezing process used in cryopreservation can injure or damage the cells due to the cytotoxicity of cryoprotecting agents (CPAs). We have previously reported a CPA-free cryopreservation method based on inkjet technology. In this method, the vitrified cells were exposed to the room temperature atmosphere during the transport of the cells using tweezers, which caused devitrification due to the increased temperature and often lowered the cell viability. In the present study, we developed an automatic thawing apparatus that transports the vitrified cells rapidly into a prewarmed medium using a spring hinge. Observations with a high-speed camera revealed that the spring hinge drops the cells into the prewarmed medium within 20 ms. All heat-transfer simulations for the apparatuses with different designs and rotation speeds showed that the cells remained below the glass-transition temperature during the transport. Finally, the apparatus was evaluated using mouse fibroblast 3T3 cells. The cell viability was improved and its reproducibility was enhanced using this apparatus. The results indicate that the combination of superflash freezing with the rapid thawing process represents a promising approach to circumvent the problems typically associated with the addition of CPAs.

中文翻译：

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在使用自动解冻装置的超快速冷冻方法中提高和可重复的细胞活力

细胞冷冻保存通过将细胞置于冷冻状态来阻止细胞的生物活性，并且可用于保存细胞而无需传代，这会导致污染和遗传漂移。然而，由于冷冻保护剂 (CPA) 的细胞毒性，冷冻保存中使用的冷冻过程可能会伤害或损坏细胞。我们之前曾报道过一种基于喷墨技术的无 CPA 冷冻保存方法。在这种方法中，在使用镊子运输细胞期间，玻璃化的细胞暴露在室温气氛中，这会由于温度升高而导致失透，并且通常会降低细胞活力。在本研究中，我们开发了一种自动解冻装置，使用弹簧铰链将玻璃化细胞快速输送到预热的培养基中。高速相机的观察表明，弹簧铰链在 20 毫秒内将细胞放入预热的培养基中。具有不同设计和旋转速度的设备的所有传热模拟表明，电池在运输过程中保持在玻璃化转变温度以下。最后，使用小鼠成纤维细胞 3T3 细胞评估该装置。使用该装置提高了细胞活力并增强了其再现性。结果表明，超快速冷冻与快速解冻过程的结合代表了一种有前途的方法来规避通常与添加 CPA 相关的问题。具有不同设计和旋转速度的设备的所有传热模拟表明，电池在运输过程中保持在玻璃化转变温度以下。最后，使用小鼠成纤维细胞 3T3 细胞评估该装置。使用该装置提高了细胞活力并增强了其再现性。结果表明，超快速冷冻与快速解冻过程的结合代表了一种有前途的方法来规避通常与添加 CPA 相关的问题。具有不同设计和旋转速度的设备的所有传热模拟表明，电池在运输过程中保持在玻璃化转变温度以下。最后，使用小鼠成纤维细胞 3T3 细胞评估该装置。使用该装置提高了细胞活力并增强了其再现性。结果表明，超快速冷冻与快速解冻过程的结合代表了一种有前途的方法来规避通常与添加 CPA 相关的问题。