Research on the Cryopreservation System of Controlled Rapid Cooling and Rewarming of Cellular Micro-droplets

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Research on the Cryopreservation System of Controlled Rapid Cooling and Rewarming of Cellular Micro-droplets

更新时间：2025-02-28

- Research on the Cryopreservation System of Controlled Rapid Cooling and Rewarming of Cellular Micro-droplets
- Journal of refrigeration (2025)
- 作者机构：
  
  上海理工大学,上海市上海市200082,中国
- 作者简介：
- 基金信息：
  
  the National Natural Science Foundation of China(52076140)
- DOI：
  CLC：
- Received：27 November 2024，
  
  Revised：22 December 2024，
  
  Accepted：22 January 2025，
- 稿件说明：
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Research on the Cryopreservation System of Controlled Rapid Cooling and Rewarming of Cellular Micro-droplets[J/OL]. Journal of refrigeration, 2025.
DOI：

Research on the Cryopreservation System of Controlled Rapid Cooling and Rewarming of Cellular Micro-droplets[J/OL]. Journal of refrigeration, 2025. DOI：

摘要

细胞微液滴的玻璃化保存方法在低温保存领域受到广泛关注，但微液滴玻璃化过程还存在降温/复温过程不可控等局限性。本文采用移动滑台升降，结合焦耳热复温方法搭建用于细胞微液滴玻璃化过程可控快速降温及复温的冻存系统，该系统通过调节浸入液氮的速度实现快速、可控降温，并通过控制焦耳热的加热时间、电流大小实现高速率复温过程，实现微液滴的玻璃化过程并显著避免了液滴复温过程的反玻璃化对细胞的损伤。结果表明：本系统可控制降温速率可以达到1.8×10

℃/min，能够实现细胞微液滴较低浓度保护剂下的玻璃化保存；复温速率控制可以达到4.0×10

℃/min，能有效避免较低浓度保护剂复温过程反玻璃化及冰晶再生长的发生；通过微液滴A549细胞低温保存效果，证实了本系统冷冻复苏后的存活率显著高于传统麦管保存。本文研究工作有望为细胞微液滴自动玻璃化保存及复温方法提供新的解决方案。

Abstract

The vitrification preservation method of cell microdroplets has received extensive attention in the field of cryopreservation. However

the vitrification process of microdroplets still has limitations such as uncontrollable cooling/rewarming processes. In this paper

a cryopreservation system for controllable rapid cooling and rewarming during the vitrification process of cell microdroplets was constructed by combining the lifting of a moving slide table with the Joule heating rewarming method. This system achieves rapid and controllable cooling by adjusting the speed of immersion in liquid nitrogen

and realizes a high-rate rewarming process by controlling the heating time and current intensity of Joule heating

thus realizing the vitrification process of microdroplets and significantly avoiding the damage to cells caused by devitrification during the droplet rewarming process. The results show that the cooling rate controlled by this system can reach 1.8 × 10⁴ °C/min

enabling the vitrification preservation of cell microdroplets with a relatively low concentration of cryoprotectant. The rewarming rate control can reach 4.0 × 10⁴ °C/min

which can effectively avoid the occurrence of devitrification and ice crystal regrowth during the rewarming process of the relatively low concentration of cryoprotectant. Through the cryopreservation effect of A549 cells in microdroplets

it has been confirmed that the survival rate after cryopreservation and resuscitation of this system is significantly higher than that of the traditional straw preservation. The research work in this paper is expected to provide new solutions for the automatic vitrification preservation and rewarming methods of cell microdroplets.

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references

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