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Mechanism and Optimization Strategy of Cryopreservation Damage in Microcarrier-Hepatocyte Complex
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Binjiang Hao, Zengying Yu, Baolin Liu
Journal of Refrigeration | 2025, 46(4) : 156 - 162
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Journal of Refrigeration | 2025, 46(4): 156-162
Mechanism and Optimization Strategy of Cryopreservation Damage in Microcarrier-Hepatocyte Complex
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Binjiang Hao, Zengying Yu, Baolin Liu
Affiliations
  • Institute of Biothermal and Technology, University of Shanghai for Science and Technology, Shanghai, 200093, China
Published: 2025-08-16 doi: 10.12465/j.issn.0253-4339.2025.04.156
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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 (Me2SO) 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 Me2SO 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.

cryopreservation  /  hepatocyte-microcarrier complex  /  freeze protectants  /  thermal expansion damage
Binjiang Hao, Zengying Yu, Baolin Liu. 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
Funding
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  • National Natural Science Foundation of China(51776130)
Appendix
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Year 2025 volume 46 Issue 4
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Article Info
doi: 10.12465/j.issn.0253-4339.2025.04.156
  • Receive Date：2024-08-04
  • Online Date：2026-03-13
  • Published：2025-08-16
Article Data
Affiliations
History
  • Received：2024-08-04
  • Revised：2024-08-29
  • Accepted：2024-09-26
Funding
National Natural Science Foundation of China(51776130)
Affiliations
    Institute of Biothermal and Technology, University of Shanghai for Science and Technology, Shanghai, 200093, China

Corresponding:

Liu Baolin, male, professor, Institute of Biothermal Science and Technology, University of Shanghai for Science and Technology, 86-13636524955, E-mail: . Research fields: the cryopreservation of cells, tissues and organs, food and drug refrigeration and drying technology.
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小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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