Rate-­‐controlled Cryopreservation and Thawing of Mammalian Cells

Thompson, Maria; Nemits, Michelle; Ehrhardt, Rolf O.

doi:10.1038/protex.2011.224

Cited by 7 publications

(8 citation statements)

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“…18,20,25,40–42 These solutions are non-toxic and easily soluble and are thought to prevent such injury via several proposed mechanisms. 18,20,22,25,40–44 They can partially solubilize the cell membrane so that it is less prone to puncture and interrupt the lattice of the ice, so that fewer intracellular crystals form. Cryoprotectants also increase the solute concentration in cells thus lowering the glass transition temperature (for water is −135C) of a solution below which molecular movement ceases and all biological activity is suspended thus preventing actual freezing and the cytosol maintains some flexibility in a glassy phase.…”

Section: Discussionmentioning

confidence: 99%

“…In order to prevent such freezing injury various cryoprotectants such as glycerol, ethanediol, propanediol and dimethyl sulfoxide (DMSO) have been used for stable freezing and long-term preservation of established cell lines and tissues. 22–27 Cryopreservation of primary patient tumor and subsequent PDX has been investigated previously, but the efficacy of reanimation has typically been low, or not reported. 23,24,28,29 Cryopreservation of composite tissues, such as PDX tumors, is more complex than for homogenous cell lines as a three-dimensional architecture needs to be maintained during both freezing and thawing.…”

Section: Introductionmentioning

confidence: 99%

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Patient-derived xenograft cryopreservation and reanimation outcomes are dependent on cryoprotectant type

Ivanics

Bergquist

Liu

et al. 2018

Laboratory Investigation

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Introduction Patient-derived xenografts (PDX) are being increasingly utilized in preclinical oncologic research. Maintaining large colonies of early generation tumor-bearing mice is impractical and cost-prohibitive. Optimal methods for efficient long-term cryopreservation and subsequent reanimation of PDX tumors are critical to any viable PDX program. We sought to compare the performance of “Standard” and “Specialized” cryoprotectant media on various cryopreservation and reanimation outcomes in PDX tumors. Standard (10% DMSO media) and Specialized (Cryostor®) media were compared between overall and matched PDX tumors. Primary outcome was reanimation engraftment efficiency (REE). Secondary outcomes included time-to-tumor-formation (TTF), time-to-harvest (TTH), and potential loss of unique PDX lines. Overall 57 unique PDX tumors underwent 484 reanimation engraftment attempts after previous cryopreservation. There were 10 unique PDX tumors cryopreserved with Standard (71 attempts), 40 with Specialized (272 attempts), and 7 with both (141 attempts). Median frozen time of reanimated tumors was 29 weeks (max.177). Tumor pathology, original primary PDX growth rates, frozen storage times, and number of implantations per PDX model were similar between cryoprotectant groups. Specialized media resulted in superior REE (overall: 82% vs. 39%, p<0.0001; matched: 97% vs. 36%, p<0.0001; >52 weeks cryostorage: 59% vs. 9%, p<0.0001), shorter TTF (overall 24 vs. 54 days, p=0.0051; matched 18 vs. 53 days, p=0.0013) and shorter TTH (overall: 64 vs. 89 days, p=0.009; matched: 47 vs. 88 days, p=0.0005) compared to Standard. Specialized media demonstrated improved REE with extended duration cryostorage (p=0.048) compared to Standard. Potential loss of unique PDX lines was lower with Specialized media (9% vs. 35%, p=0.017). In conclusion, cryopreservation with a specialized cryoprotectant appears superior to traditional laboratory-based media and can be performed with reliable reanimation even after extended cryostorage.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Patient-derived xenograft cryopreservation and reanimation outcomes are dependent on cryoprotectant type

Ivanics

Bergquist

Liu

et al. 2018

Laboratory Investigation

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“…In SF, a phase transition occurs from liquid to solid on temperatures below freezing point. Slow-cooling protocols involve the use of <1.0 M of cryoprotective agents (CPAs), such as glycerol or dimethyl sulphoxide (DMSO), which have minimal toxicity at lower temperatures with the use of a high-cost controlled rate freezer or a benchtop portable freezing container [21].…”

Section: Slow Freezingmentioning

confidence: 99%

Cryopreservation Methods and Frontiers in the Art of Freezing Life in Animal Models

Aljaser¹

2022

Veterinary Medicine and Science

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The development in cryobiology in animal breeding had revolutionized the field of reproductive medicine. The main objective to preserve animal germplasm stems from variety of reasons such as conservation of endangered animal species, animal diversity, and an increased demand of animal models and/or genetically modified animals for research involving animal and human diseases. Cryopreservation has emerged as promising technique for fertility preservation and assisted reproduction techniques (ART) for production of animal breeds and genetically engineered animal species for research. Slow rate freezing and rapid freezing/vitrification are the two main methods of cryopreservation. Slow freezing is characterized by the phase transition (liquid turning into solid) when reducing the temperature below freezing point. Vitrification, on the other hand, is a phenomenon in which liquid solidifies without the formation of ice crystals, thus the process is referred to as a glass transition or ice-free cryopreservation. The vitrification protocol applies high concentrations of cryoprotective agents (CPA) used to avoid cryoinjury. This chapter provides a brief overview of fundamentals of cryopreservation and established methods adopted in cryopreservation. Strategies involved in cryopreserving germ cells (sperm and egg freezing) are included in this chapter. Last section describes the frontiers and advancement of cryopreservation in some of the important animal models like rodents (mouse and rats) and in few large animals (sheep, cow etc).

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“…There are two methods in the cryopreservation of embryo: vitrification and programmed slow freezing (Arav 2014). In literature, some considerations were identified in the programmed slow freezing, Thompson et al (2011) indicated that subjecting embryos to a rate of 1 °C min -1 is considered a typical cooling rate for mammalian embryos. Despite the huge costs, equipment, and multiple steps of slow freezing, it has been observed a decrease in both survival and implantation rates (Bromfield et al 2009).…”

Section: Introductionmentioning

confidence: 99%

Efficiency of Dimethyl Sulphoxide and Ethylene Glycol on Subsequent Development of Vitrified Awassi Sheep Embryos

Mardenli

Al-Kerwi

Hassooni

2020

JITV

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The use of cryoprotectants in vitrification would reduce the critical damages to the embryos, thus increase the survival rates. This research was conducted in the laboratory of reproductive biotechnology at the faculty of Agriculture of Aleppo University. The study aimed to evaluate the viability and survivability of early Syrian Awassi embryos under the influence of dimethyl sulphoxide (DMSO) and ethylene glycol (EG) following vitrification. Embryos were vitrified in three solutions of cryoprotectants (A: DMSO (3 ml), B: EG (3 ml), and C which was composed of a combination of DMSO (1.5 ml) and EG (1.5 ml)). After thawing, embryos that had been vitrified in C solution achieved the highest rates of cleavage (P< 0.01) comparing with A and B solutions for 2-16 cell stage (50.00% Vs 30.77% and 36.36%), morula (9.00% Vs 44.44% and 40.00%) and blastocyst stage embryos (92.86% Vs 58.33% and 50.00%) respectively. Down to the hatching blastocyst stage, 2-16 cell stage vitrified embryos in C solution achieved an encouraging rate comparing with A and B solutions (39.20% Vs23.08% and 22.73% respectively). The rates of arrested embryos decreased significantly (P< 0.05) after thawing across the three solutions especially the morula and blastocyst stage (0.00 and 3.70% respectively) (C solution). No significant differences were observed in the three types of embryos across all stages and solutions despite the large range among these rates. Given the apparent benefit of the participatory effect of cytoprotectants, it is advised to use a mixture of DMSO and EG (1:1) in vitrification of ovine embryos.

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Rate-‐controlled Cryopreservation and Thawing of Mammalian Cells

Cited by 7 publications

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Patient-derived xenograft cryopreservation and reanimation outcomes are dependent on cryoprotectant type

Patient-derived xenograft cryopreservation and reanimation outcomes are dependent on cryoprotectant type

Cryopreservation Methods and Frontiers in the Art of Freezing Life in Animal Models

Efficiency of Dimethyl Sulphoxide and Ethylene Glycol on Subsequent Development of Vitrified Awassi Sheep Embryos

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Rate-­‐controlled Cryopreservation and Thawing of Mammalian Cells

Cited by 7 publications

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Patient-derived xenograft cryopreservation and reanimation outcomes are dependent on cryoprotectant type

Patient-derived xenograft cryopreservation and reanimation outcomes are dependent on cryoprotectant type

Cryopreservation Methods and Frontiers in the Art of Freezing Life in Animal Models

Efficiency of Dimethyl Sulphoxide and Ethylene Glycol on Subsequent Development of Vitrified Awassi Sheep Embryos

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Rate-‐controlled Cryopreservation and Thawing of Mammalian Cells