ArticlePloS one2018
Directional freezing for the cryopreservation of adherent mammalian cells on a substrate.
Article in PloS one, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- Bottom-Up Ice Growth Geometry Attenuates Shear Stress and Improves the Cryopreservation of Hematopoietic Stem/Progenitor Cells Under Low DMSO Concentrations.Biotechnology and bioengineering · 2026Article
- Cryopreservation Strategies to Improve Access to Organ Transplantation.Transplantation · 2026Review
- Thermodynamic Principles of Emerging Cryopreservation Technologies.Methods in molecular biology (Clifton, N.J.) · 2026Review
- Supercooled preservation of cultured primary rat hepatocyte monolayers.Frontiers in bioengineering and biotechnology · 2024Article
- Cryopreservation of assay-ready hepatocyte monolayers by chemically-induced ice nucleation: preservation of hepatic function and hepatotoxicity screening capabilities.Biomaterials science · 2023Article
- Effects of human placenta cryopreservation on molecular characteristics of placental mesenchymal stromal cells.Frontiers in bioengineering and biotechnology · 2023Article
- Article
- Assay-ready Cryopreserved Cell Monolayers Enabled by Macromolecular Cryoprotectants.Biomacromolecules · 2022Article
- Cryopreservation by Directional Freezing and Vitrification Focusing on Large Tissues and Organs.Cells · 2022Review
- Article
- Winter is coming: the future of cryopreservation.BMC biology · 2021Review
- Xeno-free cryopreservation of adherent retinal pigmented epithelium yields viable and functional cells in vitro and in vivo.Scientific reports · 2021Article
- Cryopreservation of human cerebral microvascular endothelial cells and astrocytes in suspension and monolayers.PloS one · 2021Article
- Preservation solutions for attenuation of ischemia-reperfusion injury in vascularized composite allotransplantation.SAGE open medicine · 2021Article
- Extracellular Antifreeze Protein Significantly Enhances the Cryopreservation of Cell Monolayers.Biomacromolecules · 2019Article
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Successfully cryopreserving cells adhered to a substrate would facilitate the growth of a vital confluent cell culture after thawing while dramatically shortening the post-thaw culturing time. Herein we propose a controlled slow cooling method combining initial directional freezing followed by gradual cooling down to -80°C for robust preservation of cell monolayers adherent to a substrate. Using computer controlled cryostages we examined the effect of cooling rates and dimethylsulfoxide (DMSO) concentration on cell survival and established an optimal cryopreservation protocol. Experimental results show the highest post-thawing viability for directional ice growth at a speed of 30 μm/sec (equivalent to freezing rate of 3.8°C/min), followed by gradual cooling of the sample with decreasing rate of 0.5°C/min. Efficient cryopreservation of three widely used epithelial cell lines: IEC-18, HeLa, and Caco-2, provides proof-of-concept support for this new freezing protocol applied to adherent cells. This method is highly reproducible, significantly increases the post-thaw cell viability and can be readily applied for cryopreservation of cellular cultures in microfluidic devices.
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Registered trials
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