2. Academic Validation
  3. Unveiling the protective role of recombinant snow flea antifreeze protein expressed by Komagataella phaffii in cryopreservation of EA.hy926 cells

Unveiling the protective role of recombinant snow flea antifreeze protein expressed by Komagataella phaffii in cryopreservation of EA.hy926 cells

  • Int J Biol Macromol. 2025 May;310(Pt 2):143295. doi: 10.1016/j.ijbiomac.2025.143295.
Yishan Fu 1,  Bo Zeng 2,  Yangyang Li 1,  Zihe Lin 1,  Shaoxiong Shen 2,  Song Liu 3,  Jian Chen 4
Affiliations

Affiliations

  • 1 Science Center for Future Foods, Jiangnan University, Wuxi 214122, China; School of Biotechnology, Jiangnan University, Wuxi 214122, China.
  • 2 Science Center for Future Foods, Jiangnan University, Wuxi 214122, China; School of Biotechnology, Jiangnan University, Wuxi 214122, China; JiaXing Institute of Future Food, Jiaxing, Zhejiang 314000, China.
  • 3 Science Center for Future Foods, Jiangnan University, Wuxi 214122, China; School of Biotechnology, Jiangnan University, Wuxi 214122, China. Electronic address: liusong@jiangnan.edu.cn.
  • 4 Science Center for Future Foods, Jiangnan University, Wuxi 214122, China; School of Biotechnology, Jiangnan University, Wuxi 214122, China. Electronic address: jchen@jiangnan.edu.cn.
PMID: 40258546 DOI: 10.1016/j.ijbiomac.2025.143295
Abstract

Cryopreservation is pivotal for preserving the viability and functionality of cells, tissues, and organs, which is crucial for medical research and therapeutic applications. In this study, recombinant snow flea antifreeze protein (rSfAFP) was produced using Komagataella phaffii GS115 and evaluated for cryopreservation of human umbilical vein endothelial cells (EA.hy926). rSfAFP demonstrated a significant thermal hysteresis activity of 2.5 °C at 5 mg/mL and effectively suppressed ice recrystallization. At a concentration of 1 mg/mL, rSfAFP maintained 67.6 % cell recovery rate post-cryopreservation, a mere 15 % decrease compared to cells cryopreserved using 8 % dimethyl sulfoxide. Proteomic analysis revealed that rSfAFP enhances cell survival by preserving cellular membrane integrity, reducing Reactive Oxygen Species, and promoting protein synthesis, similar to dimethyl sulfoxide but with distinct protein expression patterns. This research underscores the potential application of rSfAFP in cell cryopreservation and provides insights into the mechanisms by which antifreeze proteins enhance cell recovery rate after thawing.

Keywords

Antifreeze protein; Cell recovery rate; Cryopreservation; Snow flea; Thermal hysteresis.

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