Cryopreservation of Liver-Cell Spheroids with Macromolecular Cryoprotectants

Bissoyi, Akalabya1; Tomas, Ruben M. F.1; Gao, Yanan2,3; Guo, Qiongyu4; Gibson, Matthew I.1,2

2023

10.1021/acsami.2c18288

ACS Applied Materials & Interfaces   Impact Factor & Quartile

1944-8244

1944-8252

Spheroids are a powerful tool for basic research and to reduce or replace in vivo (animal) studies but are not routinely banked nor shared. Here, we report the successful cryopreservation of hepatocyte spheroids using macromolecular (polyampholyte) cryoprotectants supplemented into dimethyl sulfoxide (DMSO) solutions. We demonstrate that a polyampholyte significantly increases post-thaw recovery, minimizes membrane damage related to cryo-injury, and remains in the extracellular space making it simple to remove post-thaw. In a model toxicology challenge, the thawed spheroids matched the performance of fresh spheroids. F-actin staining showed that DMSO-only cryopreserved samples had reduced actin polymerization, which the polyampholyte rescued, potentially linked to intracellular ice formation. This work may facilitate access to off-the-shelf and ready-to-use frozen spheroids, without the need for in-house culturing. Readily accessible 3-D cell models may also reduce the number of in vivo experiments.

macromolecular cryoprotectants polymers spheroids cryopreservation cell-based assays macromolecules ice

SCI

English

Others

European Research Council (ERC) under European Union["866056","899872"] ; Royal Society for an Industry Fellowship[191037] ; University of Warwick[BB/W510907/1] ; InnovateUK[10004515] ; National Natural Science Foundation of China[81971764]

Science & Technology - Other Topics ; Materials Science

Nanoscience & Nanotechnology ; Materials Science, Multidisciplinary

WOS:000914744600001

AMER CHEMICAL SOC

Web of Science

1.Univ Warwick, Warwick Med Sch, Div Biomed Sci, Coventry CV4 7AL, England
2.Univ Warwick, Dept Chem, Coventry CV4 7AL, England
3.Southern Univ Sci & Technol, Dept Biomed Engn, Shenzhen 518055, Guangdong, Peoples R China
4.Southern Univ Sci & Technol, Dept Biomed Engn, Shenzhen 518055, Guangdong, Peoples R China

Bissoyi, Akalabya,Tomas, Ruben M. F.,Gao, Yanan,et al. Cryopreservation of Liver-Cell Spheroids with Macromolecular Cryoprotectants[J]. ACS Applied Materials & Interfaces,2023.

Bissoyi, Akalabya,Tomas, Ruben M. F.,Gao, Yanan,Guo, Qiongyu,&Gibson, Matthew I..(2023).Cryopreservation of Liver-Cell Spheroids with Macromolecular Cryoprotectants.ACS Applied Materials & Interfaces.

Bissoyi, Akalabya,et al."Cryopreservation of Liver-Cell Spheroids with Macromolecular Cryoprotectants".ACS Applied Materials & Interfaces (2023).