Chemically Induced Extracellular Ice Nucleation Reduces Intracellular Ice Formation Enabling 2D and 3D Cellular Cryopreservation

Murray，Kathryn A.1,2; Gao，Yanan1,3; Griffiths，Christopher A.4; Kinney，Nina L.H.1; Guo，Qiongyu3; Gibson，Matthew I.1,2; Whale，Thomas F.1

2023

10.1021/jacsau.3c00056

JACS Au   影响因子和分区

2691-3704

3D cell assemblies such as spheroids reproduce the in vivo state more accurately than traditional 2D cell monolayers and are emerging as tools to reduce or replace animal testing. Current cryopreservation methods are not optimized for complex cell models, hence they are not easily banked and not as widely used as 2D models. Here we use soluble ice nucleating polysaccharides to nucleate extracellular ice and dramatically improve spheroid cryopreservation outcomes. This protects the cells beyond using DMSO alone, and with the major advantage that the nucleators function extracellularly and hence do not need to permeate the 3D cell models. Critical comparison of suspension, 2D and 3D cryopreservation outcomes demonstrated that warm-temperature ice nucleation reduces the formation of (fatal) intracellular ice, and in the case of 2/3D models this reduces propagation of ice between adjacent cells. This demonstrates that extracellular chemical nucleators could revolutionize the banking and deployment of advanced cell models.

3D cell assemblies chemical control of ice formation Cryopreservation ice nucleation polysaccharides

ESCI

英语

其他

European Research Council (ERC) under the European Union[866056] ; Royal Society[191037] ; University of Warwick["WT-219429/Z/19/Z","ECF-2018-127"] ; Natural Environment Research Council[NE/S007350/1] ; National Natural Science Foundation of China[81971764]

Chemistry

Chemistry, Multidisciplinary

WOS:000980970900001

AMER CHEMICAL SOC

2-s2.0-85156225292

Scopus

1.Department of Chemistry,University of Warwick,Coventry,Gibbet Hill Road,CV4 7AL,United Kingdom
2.Division of Biomedical Sciences,Warwick Medical School,University of Warwick,Coventry,Gibbet Hill Road,CV4 7AL,United Kingdom
3.Department of Biomedical Engineering,Southern University of Science and Technology,Shenzhen,Guangdong,518055,China
4.Department of Aquatic Resources,Institute of Marine Research,Swedish University of Agricultural Sciences,Lysekil,Turistgatan 5,453 30,Sweden

Murray，Kathryn A.,Gao，Yanan,Griffiths，Christopher A.,et al. Chemically Induced Extracellular Ice Nucleation Reduces Intracellular Ice Formation Enabling 2D and 3D Cellular Cryopreservation[J]. JACS Au,2023,3(5).

Murray，Kathryn A..,Gao，Yanan.,Griffiths，Christopher A..,Kinney，Nina L.H..,Guo，Qiongyu.,...&Whale，Thomas F..(2023).Chemically Induced Extracellular Ice Nucleation Reduces Intracellular Ice Formation Enabling 2D and 3D Cellular Cryopreservation.JACS Au,3(5).

Murray，Kathryn A.,et al."Chemically Induced Extracellular Ice Nucleation Reduces Intracellular Ice Formation Enabling 2D and 3D Cellular Cryopreservation".JACS Au 3.5(2023).