Cryopreservation and Rapid Recovery of Differentiated Intestinal Epithelial Barrier Cells at Complex Transwell Interfaces Is Enabled by Chemically Induced Ice Nucleation

Bissoyi, Akalabya1,2; Gao, Yanan3,4; Tomas, Ruben M. F.5; Kinney, Nina L. H.3,6; Whale, Thomas F.3,7; Guo, Qiongyu4; Gibson, Matthew I.1,2,3,8

2024-04-26

10.1021/acsami.4c03931

ACS APPLIED MATERIALS & INTERFACES   影响因子和分区

1944-8244

1944-8252

Cell-based models, such as organ-on-chips, can replace and inform in vivo (animal) studies for drug discovery, toxicology, and biomedical science, but most cannot be banked "ready to use" as they do not survive conventional cryopreservation with DMSO alone. Here, we demonstrate how macromolecular ice nucleators enable the successful cryopreservation of epithelial intestinal models supported upon the interface of transwells, allowing recovery of function in just 7 days post-thaw directly from the freezer, compared to 21 days from conventional suspension cryopreservation. Caco-2 cells and Caco-2/HT29-MTX cocultures are cryopreserved on transwell inserts, with chemically induced ice nucleation at warmer temperatures resulting in increased cell viability but crucially retaining the complex cellular adhesion on the transwell insert interfaces, which other cryoprotectants do not. Trans-epithelial electrical resistance measurements, confocal microscopy, histology, and whole-cell proteomics demonstrated the rapid recovery of differentiated cell function, including the formation of tight junctions. Lucifer yellow permeability assays confirmed that the barrier functions of the cells were intact. This work will help solve the long-standing problem of transwell tissue barrier model storage, facilitating access to advanced predictive cellular models. This is underpinned by precise control of the nucleation temperature, addressing a crucial biophysical mode of damage.

cryopreservation nucleation Caco-2 cells toxicology ice

SCI ; EI

英语

其他

European Research Council (ERC) under the European Union["866056","899872"] ; Royal Society for an Industry Fellowship[191037] ; InnovateUK[10004515] ; National Natural Science Foundation of China[81971764] ; Leverhulme Trust[ECF-2018-127] ; Natural Environment Research Council[NE/S007350/1]

Science & Technology - Other Topics ; Materials Science

Nanoscience & Nanotechnology ; Materials Science, Multidisciplinary

WOS:001227717800001

AMER CHEMICAL SOC

Web of Science

1.Univ Manchester, Dept Chem, Manchester M13 9PL, England
2.Univ Manchester, Manchester Inst Biotechnol, Manchester M1 7DN, England
3.Univ Warwick, Dept Chem, Coventry CV4 7AL, England
4.Southern Univ Sci & Technol, Dept Biomed Engn, Shenzhen 518055, Guangdong, Peoples R China
5.Univ Warwick Sci Pk, Venture Ctr, Univ Warwick Sci Pk, Coventry CV4 7EZ, England
6.Royal Bot Gardens, Ardingly RH17 6TN, W Sussex, England
7.Univ Leeds, Sch Earth & Environm, Leeds LS2 9JT, England
8.Univ Warwick, Warwick Med Sch, Div Biomed Sci, Coventry CV4 7AL, England

Bissoyi, Akalabya,Gao, Yanan,Tomas, Ruben M. F.,et al. Cryopreservation and Rapid Recovery of Differentiated Intestinal Epithelial Barrier Cells at Complex Transwell Interfaces Is Enabled by Chemically Induced Ice Nucleation[J]. ACS APPLIED MATERIALS & INTERFACES,2024,16(18).

Bissoyi, Akalabya.,Gao, Yanan.,Tomas, Ruben M. F..,Kinney, Nina L. H..,Whale, Thomas F..,...&Gibson, Matthew I..(2024).Cryopreservation and Rapid Recovery of Differentiated Intestinal Epithelial Barrier Cells at Complex Transwell Interfaces Is Enabled by Chemically Induced Ice Nucleation.ACS APPLIED MATERIALS & INTERFACES,16(18).

Bissoyi, Akalabya,et al."Cryopreservation and Rapid Recovery of Differentiated Intestinal Epithelial Barrier Cells at Complex Transwell Interfaces Is Enabled by Chemically Induced Ice Nucleation".ACS APPLIED MATERIALS & INTERFACES 16.18(2024).