Search results
Charles J. Hunt (April 8, 1881 – February 3, 1976) was an American film editor and director. He also worked at various times as an actor, production manager and associate producer.
Charles John Hunt. Cryopreservation is the application of low temperatures to preserve the structural and functional integrity of cells and tissues. Conventional cooling protocols allow ice to...
Cryopreservation is the application of low temperatures to preserve the structural and functional integrity of cells and tissues. Conventional cooling protocols allow ice to form and solute concentrations to rise during the cryopreservation process.
- Charles J. Hunt
- 2017
Author. Charles J Hunt 1. Affiliation. 1 Sawston, United Kingdom. PMID: 31244583. PMCID: PMC6558338. DOI: 10.1159/000497289. Abstract. The commercial and clinical development of cellular therapy products will invariably require cryopreservation and frozen storage of cellular starting materials, intermediates and/or final product.
- Charles J Hunt
- 2019
- Keywords
- Abstract
- Introduction
- Choice of CPA
- Automated Filling Systems
- Self-Formulated CPAs
- Other cryoprotectant solutions
- Supercooling: An as Yet Uncontrolled Variable
- Disaster Recovery
- Thawing and Elution of CPA
- Elution of CPA
- Conclusions
- Disclosure Statement
Cryopreservation · Cryoprotectants · Freezing · Low-temperature storage · Thawing · Stem cells · Cell therapy
The commercial and clinical development of cellular therapy products will invariably require cryopreservation and frozen storage of cellular starting materials, intermediates and/or final product. Optimising cryopreservation is as important as optimisation of the cell culture process in obtaining maxi-mum yield and a consistent end-product. Subopti...
Regenerative medicine has been defined as the re-placement or regeneration of human cells, tissue or or-gans to restore or establish normal function [1]. It encom-passes a wide range of therapeutic modalities from organ and tissue transplantation to sophisticated tissue-engi-neered scaffolds and cellular therapies, as well as more traditional treat...
CPAs protect cells predominantly from the damaging effects of freezing at slow cooling rates (i.e., those gener-ally achievable using commercially available passive cool-ing devices [PCDs] and controlled-rate freezers [CRFs]). When added in sufficient concentration, they depress the freezing point of the solution, reducing the amount of ice formed ...
Once exposed to the CPA, reducing the time taken be-fore commencing the freezing process will help reduce unwanted cytotoxic effects. With medium to large batch sizes the time taken to manually aliquot the cell suspen-sion into vials can be significant. Moreover, maintaining a low temperature within a cleanroom environment is of-ten difficult. The ...
The availability of CE-marked, sterile CPAs such as DMSO (Table 1) and appropriate vehicle solutions means that self-formulated cryoprotectant solutions can be pre-pared in-house. The use of self-formulated CPAs, the prac-tice generally adopted in a research environment, while po-tentially advantageous during the optimisation phase of the cryoprote...
a The list is not intended to be definitive. All information is taken from supplier websites. Only CPAs indicated as intended for use in therapeutic applications are listed. respond to the rate of change in temperature by water ef-flux, while rapid cooling can be loosely defined as that where IIF predominates. This emphasises the contextual nature...
Of all the events occurring during cryopreservation, supercooling is the least controlled [53]. Supercooling (or undercooling) is the temperature difference between the equilibrium melting point of the system and the temper-ature at which nucleation occurs. The degree of super-cooling can be highly variable between samples cooled during the same co...
Off-site, secondary storage of material for disaster re-covery purposes is considered best practice and may be a regulatory requirement where material is prepared for clinical applications. The material will generally come from the master cell bank but earlier, precursor stocks may also be held for such purposes as well as archive ma-terial from pr...
As with cooling rate, warming rate can have a signifi-cant impact on cellular recovery, with cells damaged through the use of inappropriate thawing and CPA elution protocols. Rapid rewarming is important. Not only does it reduce exposure time to damaging solute concentrations as ice melts during rewarming, but it also avoids potential damage caused...
Once thawed, the cells will again be exposed to the CPA with the attendant danger of chemical toxicity if held at too high a temperature. This can be mitigated by reducing the temperature (i.e., maintain cells at around 4 ° C), using a lower CPA concentration during freezing (subject to it providing optimal cryoprotection) or elut-ing it from th...
The manufacture and delivery of cellular therapy products will invariably require cryopreservation and frozen storage of cellular starting materials, intermediates and/or final product. Optimisation of culture conditions to provide maximum yield and a consistent end-product is seen as a necessary part of process development and validation, yet ofte...
The author has no conflicts of interest to declare.
Mar 28, 2019 · This paper seeks to provide practical insights and alternative solutions to the technical challenges faced during cryopreservation of cells for use in cellular therapies. Keywords: Cryopreservation, Cryoprotectants, Freezing, Low-temperature storage, Thawing, Stem cells, Cell therapy.
TLDR. This study has shown that cooling must be controlled to -40°C before transfer to long term storage to ensure optimal cell recovery, and indicates that the cells have become sufficiently dehydrated to avoid further, significant damage when transferred into ultra-low temperature storage. Expand. 8.
