Effect of Antifreeze Glycoproteins on Organoid Survival during and after Hypothermic Storage.

Guizela Huelsz-Prince,Huib J Bakker,Konrad Meister,Jeroen S Van Zon,Arthur L Devries

doi:10.3390/biom9030110

Abstract

We study the effect of antifreeze glycoproteins (AFGPs) on the survival of organoids under hypothermic conditions. We find that the survival of organoids in cold conditions depends on their developmental stage. Mature organoids die within 24 h when being stored at 4 °C, while cystic organoids can survive up to 48 h. We find that in the presence of AFGPs, the organoid survival is prolonged up to 72 h, irrespective of their developmental stage. Fluorescence microscopy experiments reveal that the AFGPs predominately localize at the cell surface and cover the cell membranes. Our findings support a mechanism in which the positive effect of AFGPs on cell survival during hypothermic storage involves the direct interaction of AFGPs with the cell membrane. Our research highlights organoids as an attractive multicellular model system for studying the action of AFGPs that bridges the gap between single-cell and whole-organ studies.

Highlights

Hypothermic preservation is a commonly used method in which cells, tissues, or organs are maintained at low temperatures (1–10 ◦ C) for short-term storage situations that enable distant transport.During hypothermic storage, cells encounter cold stress that affects their cell physiology, metabolic activity, and regulation of ion equilibration across membranes [1]
Numerous organisms have evolved adaptive mechanisms for their survival in fluctuating cold temperatures and icy environments [3]. These mechanisms involve the production of antifreeze proteins (AFPs) and antifreeze glycoproteins (AFGPs) [3,4]
We investigated the survival rate of intestinal organoids under hypothermic conditions at time periods spanning from 24 to 120 h

Summary

Introduction

Hypothermic preservation is a commonly used method in which cells, tissues, or organs are maintained at low temperatures (1–10 ◦ C) for short-term storage situations that enable distant transport.During hypothermic storage, cells encounter cold stress that affects their cell physiology, metabolic activity, and regulation of ion equilibration across membranes [1]. Hypothermic preservation is a commonly used method in which cells, tissues, or organs are maintained at low temperatures (1–10 ◦ C) for short-term storage situations that enable distant transport. Much effort has been made in the optimization of storage solutions that minimize cold-induced damage and that increase the time interval that cells and organs can be cold-stored [2]. Numerous organisms have evolved adaptive mechanisms for their survival in fluctuating cold temperatures and icy environments [3]. These mechanisms involve the production of antifreeze proteins (AFPs) and antifreeze glycoproteins (AFGPs) [3,4]. Rubinsky et al showed that the fertility of bovine oocytes [11] could be significantly improved by the addition of fish AF(G)Ps, and that rat livers can be stabilized at low temperatures using

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Conclusion