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Cryopreservation of liver-cell spheroids with macromolecular cryoprotectants
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Bissoyi, Akalabya, Tomás, Ruben M. F., Gao, Yanan, Guo, Qiongyu and Gibson, Matthew I. (2023) Cryopreservation of liver-cell spheroids with macromolecular cryoprotectants. ACS Applied Materials & Interfaces, 15 (2). pp. 2630-2638. doi:10.1021/acsami.2c18288 ISSN 1944-8244.
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Official URL: http://dx.doi.org/10.1021/acsami.2c18288
Abstract
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.
Item Type: | Journal Article | ||||||||||||||||||||||||||||||||||||
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Subjects: | Q Science > QD Chemistry Q Science > QH Natural history |
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Chemistry | ||||||||||||||||||||||||||||||||||||
Library of Congress Subject Headings (LCSH): | Cytology, Cells -- Cryopreservation, Cryopreservation of organs, tissues, etc., Polymers, Stem cells , Biological assay , Macromolecules | ||||||||||||||||||||||||||||||||||||
Journal or Publication Title: | ACS Applied Materials & Interfaces | ||||||||||||||||||||||||||||||||||||
Publisher: | American Chemical Society | ||||||||||||||||||||||||||||||||||||
ISSN: | 1944-8244 | ||||||||||||||||||||||||||||||||||||
Official Date: | 18 January 2023 | ||||||||||||||||||||||||||||||||||||
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Volume: | 15 | ||||||||||||||||||||||||||||||||||||
Number: | 2 | ||||||||||||||||||||||||||||||||||||
Page Range: | pp. 2630-2638 | ||||||||||||||||||||||||||||||||||||
DOI: | 10.1021/acsami.2c18288 | ||||||||||||||||||||||||||||||||||||
Status: | Peer Reviewed | ||||||||||||||||||||||||||||||||||||
Publication Status: | Published | ||||||||||||||||||||||||||||||||||||
Access rights to Published version: | Open Access (Creative Commons) | ||||||||||||||||||||||||||||||||||||
Date of first compliant deposit: | 12 January 2023 | ||||||||||||||||||||||||||||||||||||
Date of first compliant Open Access: | 13 January 2023 | ||||||||||||||||||||||||||||||||||||
RIOXX Funder/Project Grant: |
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