ArticleCryobiology2020
Cryopreservation of primary cultures of mammalian somatic cells in 96-well plates benefits from control of ice nucleation.
Article in Cryobiology, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed, 55 citations in OpenAlex.
- A High-Throughput Platform for Measuring and Predicting Vitrification Behavior in Multicomponent Aqueous Solutions.ACS applied materials & interfaces · 2026Article
- A high throughput platform for measuring and predicting vitrification behavior in multicomponent aqueous solutions.bioRxiv : the preprint server for biology · 2026Article
- In Situ Growth, Etching, and Charging of Nanoscale Water Ice Under Fast Electron Irradiation in Environmental TEM.Nanomaterials (Basel, Switzerland) · 2025Article
- Induced Extracellular Ice Nucleation Protects Cocultured Spheroid Interior and Exterior during Cryopreservation.ACS biomaterials science & engineering · 2025Article
- Microfluidics for the biological analysis of atmospheric ice-nucleating particles: Perspectives and challenges.Biomicrofluidics · 2025Review
- Effect of low-frequency assisted ultrasonic on cryopreservation of L-02 hepatocyte cells.Frontiers in cell and developmental biology · 2025Article
- Cryopreserved Kidney Epithelial (Vero) Cell Monolayers for Rapid Viral Quantification, Enabled by a Combination of Macromolecular Cryoprotectants.Biomacromolecules · 2024Article
- Successful expansion and cryopreservation of human natural killer cell line NK-92 for clinical manufacturing.PloS one · 2024Article
- 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
- Post-thaw application of ROCK-inhibitors increases cryopreserved T-cell yield.RSC medicinal chemistry · 2023Article
- Proline-conditioning and chemically-programmed ice nucleation protects spheroids during cryopreservation.Chemical communications (Cambridge, England) · 2023Article
- Article
- A highly active mineral-based ice nucleating agent supportsJournal of the Royal Society, Interface · 2023Article
- Cryopreservation of Liver-Cell Spheroids with Macromolecular Cryoprotectants.ACS applied materials & interfaces · 2023Article
- Assay-ready Cryopreserved Cell Monolayers Enabled by Macromolecular Cryoprotectants.Biomacromolecules · 2022Article
- Pollen derived macromolecules serve as a new class of ice-nucleating cryoprotectants.Scientific reports · 2022Article
- The role of structural order in heterogeneous ice nucleation.Chemical science · 2022Article
- Red Blood Cell Cryopreservation with Minimal Post-Thaw Lysis Enabled by a Synergistic Combination of a Cryoprotecting Polyampholyte with DMSO/Trehalose.Biomacromolecules · 2022Article
- Chemical approaches to cryopreservation.Nature reviews. Chemistry · 2022Review
Corrections and comments
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Authors and funding
9 authors at 2 institutions in 1 country.
Funding
Abstract
Cryopreservation of mammalian cells has to date typically been conducted in cryovials, but there are applications where cryopreservation of primary cells in multiwell plates would be advantageous. However excessive supercooling in the small volumes of liquid in each well of the multiwell plates is inevitable without intervention and tends to result in high and variable cell mortality. Here, we describe a technique for cryopreservation of adhered primary bovine granulosa cells in 96-well plates by controlled rate freezing using controlled ice nucleation. Inducing ice nucleation at warm supercooled temperatures (less than 5 °C below the melting point) during cryopreservation using a manual seeding technique significantly improved post-thaw recovery from 29.6% (SD = 8.3%) where nucleation was left uncontrolled to 57.7% (9.3%) when averaged over 8 replicate cultures (p < 0.001). Detachment of thawed cells was qualitatively observed to be more prevalent in wells which did not have ice nucleation control which suggests cryopreserved cell monolayer detachment may be a consequence of deep supercooling. Using an infra-red thermography technique we showed that many aliquots of cryoprotectant solution in 96-well plates can supercool to temperatures below -20 °C when nucleation is not controlled, and also that the freezing temperatures observed are highly variable despite stringent attempts to remove contaminants acting as nucleation sites. We conclude that successful cryopreservation of cells in 96-well plates, or any small volume format, requires control of ice nucleation.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.