Study on Cryopreservation of Peripheral Blood Mononuclear Cells by Ultrasonic Ice Seeding

Xu Qiang; Li Weijie; Peng Zhengxin; Ding Wei; Lin Chunyan; Shen Jing; Zhou Xinli

doi:10.12465/j.issn.0253-4339.2025.06.151

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Study on Cryopreservation of Peripheral Blood Mononuclear Cells by Ultrasonic Ice Seeding

更新时间：2025-12-03

- Study on Cryopreservation of Peripheral Blood Mononuclear Cells by Ultrasonic Ice Seeding
- Journal of Refrigeration Vol. 46, Issue 6, Pages: 151-159(2025)
- 作者机构：
  
  1.上海理工大学生物系统热科学研究所　上海　 200093
  2.上海细胞治疗集团有限公司　上海细胞治疗研究院　上海　 201800
- 作者简介：
  
  Zhou Xinli, female, professor, Institute of Biothermal Science, University of Shanghai for Science and Technology, 86-13817547878, E-mail: zjulily@163.com. Research fields: cryobiomedical technology.
- 基金信息：
- DOI：10.12465/j.issn.0253-4339.2025.06.151
  CLC： TB61⁺1;R318.52
- Received：09 September 2024，
  
  Revised：2024-10-14，
  
  Accepted：04 November 2024，
  
  Published：16 December 2025
- 稿件说明：
移动端阅览
Xu Qiang, Li Weijie, Peng Zhengxin, et al. Study on Cryopreservation of Peripheral Blood Mononuclear Cells by Ultrasonic Ice Seeding[J]. Journal of Refrigeration, 2025, 46(6): 151-159.
DOI：

Xu Qiang, Li Weijie, Peng Zhengxin, et al. Study on Cryopreservation of Peripheral Blood Mononuclear Cells by Ultrasonic Ice Seeding[J]. Journal of Refrigeration, 2025, 46(6): 151-159. DOI： 10.12465/j.issn.0253-4339.2025.06.151.

摘要

外周血单个核细胞（PBMCs）是重要的免疫细胞资源，但传统的慢速冷冻方法延缓了PBMCs的增殖，且导致T细胞亚群的损伤，因此急需开发新的冻存工艺，有效保护PBMCs的活性和功能。通过超声植冰的方式对PBMCs冻存工艺进行优化，使用流式细胞术对冻存后的PBMCs中的T细胞亚型进行分析。搭建了超声波植冰装置，通过超声波发生设备与程序降温仪相结合，实现了无接触式植冰。结果表明：引入超声植冰的冻存工艺对PBMCs的冻存效果优于常规慢速冷冻，PBMCs冻存后的活率和累计扩增倍数分别为94.97%和204.47，且初始T细胞（Tn）在冻存复温后占比高达18.35%。通过引入超声植冰对PBMCs冻存工艺的优化，提高了PBMCs冻存后的活率、累计增殖能力和T细胞亚型占比，为PBMCs的冻存提供了新的优化方案。

Abstract

Peripheral blood mononuclear cells (PBMCs) are important immune cells. However

traditional slow-freezing methods delay the proliferation of PBMCs and damage T-cell subsets. Therefore

there is an urgent need to develop an alternative cooling procedure that can effectively preserve the viability and function of PBMCs. In this study

we optimized the cryopreservation of PBMCs using ultrasonic ice seeding and analyzed post-cryopreservation T-cell subtypes using flow cytometry. An ultrasonic ice-seeding apparatus was constructed to achieve contactless ice seeding by combining an ultrasonic generating device and a controlled-rate freezer. The results showed that the cooling procedure involving ultrasonic ice seeding exhibited superior efficacy compared to the conventional slow-freezing approach. Following cryopreservation

the viability and cumulative proliferation of PBMCs were 94.97% and 204.47%

respectively. The proportion of naive T cells (Tn) after cryopreservation and thawing accounted for up to 18.35%. By incorporating ultrasonic ice seeding

the optimized cryopreservation procedure enhanced the post-thaw viability

cumulative proliferative capacity

and proportion of T-cell subtypes in PBMCs

providing a novel and effective approach for PBMC cryopreservation.

关键词

Keywords

references

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