微载体-肝细胞复合体冻存损伤机制及优化策略

郝彬江; 庾曾颖; 刘宝林

doi:10.12465/j.issn.0253-4339.2025.04.156

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微载体-肝细胞复合体冻存损伤机制及优化策略

更新时间：2025-08-28

- 微载体-肝细胞复合体冻存损伤机制及优化策略
- Mechanism and Optimization Strategy of Cryopreservation Damage in Microcarrier-Hepatocyte Complex
- 制冷学报 2025年46卷第4期页码：156-162
- 作者机构：
  
  上海理工大学生物系统热科学研究所　上海　200093
- 作者简介：
  
  刘宝林，男，教授，上海理工大学生物系统热科学研究所，13636524955，E-mail：blliuk@163.com。研究方向：细胞、组织和器官的低温保存、食品药品的冷冻干燥技术。
- 基金信息：
  
  国家自然科学基金(51776130)
- DOI：10.12465/j.issn.0253-4339.2025.04.156
  中图分类号： TB61⁺1;R318.52
- 收稿日期：2024-08-04，
  
  修回日期：2024-08-29，
  
  录用日期：2024-09-26，
  
  纸质出版日期：2025-08-16
- 稿件说明：
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郝彬江, 庾曾颖, 刘宝林. 微载体-肝细胞复合体冻存损伤机制及优化策略[J]. 制冷学报, 2025,46(4):156-162.

Hao Binjiang, Yu Zengying, Liu Baolin. Mechanism and Optimization Strategy of Cryopreservation Damage in Microcarrier-Hepatocyte Complex[J]. Journal of refrigeration, 2025, 46(4): 156-162.
郝彬江, 庾曾颖, 刘宝林. 微载体-肝细胞复合体冻存损伤机制及优化策略[J]. 制冷学报, 2025,46(4):156-162. DOI： 10.12465/j.issn.0253-4339.2025.04.156.

Hao Binjiang, Yu Zengying, Liu Baolin. Mechanism and Optimization Strategy of Cryopreservation Damage in Microcarrier-Hepatocyte Complex[J]. Journal of refrigeration, 2025, 46(4): 156-162. DOI： 10.12465/j.issn.0253-4339.2025.04.156.

摘要

为提高细胞的存活率和贴壁率，优化人工肝支持系统中微载体-肝细胞复合体的冻存方案，通过实验评估了不同冷冻保护剂体积分数、加载温度和方式对细胞冻存效果的影响。研究发现，使用体积分数为5%的二甲亚砜（Me

SO）在4 ℃预冷条件下，通过两步加载法，能显著减少细胞毒性和渗透损伤，保持较高的细胞存活

率和贴壁率。此外，还研究了降温速率和胞内冰形成对细胞冻存效果的影响，在-6 ℃植冰可以减轻胞内冰的损伤，提高细胞复苏后的贴壁率。在1 ℃/min降温速率下，体积分数为10%的Me

SO尽管存在较高的毒性和渗透损伤，但由于热膨胀差异最小，冻存效果最佳。研究结果可为人工肝支持系统中微载体-肝细胞复合体的冻存提供技术支撑。

Abstract

The aim of this study is to optimize the cryopreservation scheme for microcarrier hepatocyte complexes used in artificial liver support systems to improve cell survival and adhesion rates. The effects of cryoprotectant concentration

loading temperature

and method on cell viability were evaluated experimentally. It was found that cell toxicity and osmotic damage were reduced significantly

and higher cell survival and adhesion rates were maintained by using two-step loading of 5% volume fraction dimethyl sulfoxide (Me

SO) at 4 ℃. In addition

we investigated the effects of intracellular ice formation and cooling rate on cell viability and attachment. By performing ice seeding at -6 ℃

the intracellular ice damage was effectively reduced

and the adhesion rate of cells after recovery was improved. The experimental results show that a 10% volume fraction of Me

SO and a cooling rate of 1 ℃/min

despite having high toxicity and osmotic damage

have the best freezing effect due to the smallest difference in thermal expansion. This study provides important techniques for the cryopreservation of microcarrier hepatocyte complexes for artificial liver support systems.

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references

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