scholarly journals The transfer temperature from slow cooling to cryogenic storage is critical for optimal recovery of cryopreserved mammalian cells

PLoS ONE ◽  
2021 ◽  
Vol 16 (11) ◽  
pp. e0259571
Author(s):  
Peter Kilbride ◽  
Julie Meneghel ◽  
Fernanda Fonseca ◽  
John Morris

Cryopreservation is a key step for the effective delivery of many cell therapies and for the maintenance of biological materials for research. The preservation process must be carefully controlled to ensure maximum, post-thaw recovery using cooling rates slow enough to allow time for cells to cryodehydrate sufficiently to avoid lethal intracellular ice. This study focuses on determining the temperature necessary at the end of controlled slow cooling before transfer to cryogenic storage which ensures optimal recovery of the processed cell samples. Using nucleated, mammalian cell lines derived from liver (HepG2), ovary (CHO) and bone tissue (MG63) this study has shown that cooling must be controlled to -40°C before transfer to long term storage to ensure optimal cell recovery. No further advantage was seen by controlling cooling to lower temperatures. These results are consistent with collected differential scanning calorimetry data, that indicated the cells underwent an intracellular, colloidal glass transition between -49 and -59°C (Tg’i) in the presence of the cryoprotective agent dimethyl sulfoxide (DMSO). The glass forms at the point of maximum cryodehydration and no further cellular dehydration is possible. At this point the risk of lethal intracellular ice forming on transfer to ultra-low temperature storage is eliminated. In practice it may not be necessary to continue slow cooling to below this temperature as optimal recovery at -40°C indicates that the cells have become sufficiently dehydrated to avoid further, significant damage when transferred into ultra-low temperature storage.

Alloy Digest ◽  
1964 ◽  
Vol 13 (8) ◽  

Abstract USS 9% Nickel Steel was specifically developed for low-temperature storage vessels operating down to minus 320 F. It is recommended for cryogenic service. This datasheet provides information on composition, physical properties, elasticity, and tensile properties as well as fracture toughness. Filing Code: SA-166. Producer or source: United States Steel Corporation.


2006 ◽  
Vol 75 (3) ◽  
pp. 209-212 ◽  
Author(s):  
Kodthalu Seetharamaiah Shivashankara ◽  
Seiichiro Isobe ◽  
Hiroshi Horita ◽  
Makiko Takenaka ◽  
Takeo Shiina

2020 ◽  
Vol 147 ◽  
pp. 01015
Author(s):  
Dimas Fendy Pradana ◽  
Ignatius Hardaningsih ◽  
Dini Wahyu Kartika Sari

The objectives of this study were to evaluate the sperm viability of Najawa carp (Cyprinus carpio L.) in cryopreservation pre-conditions at 4°C. The design used in this study was Complete Randomized Design with 4 treatments, BSS as a control, 10% DMSO, 0,2 M Sucrose, and 5% DMSO + 0,1 M Sucrose; each consist of three replications. The parameters observed were progressive motility of fresh sperm, diluted sperm before low temperature storage, and 2 hours; 3 hours; 4 hours; 5 hours; one day; one week; a month after 4°C storage. The data were analyzed by ANOVA. The data showed that there was no significant difference between treatment (P>0.05). The best viability was 40.56% of sperm motility which survive for one week, it was achieved by 5% DMSO + 0,1 M Sucrose.


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