Cryopretectants and Their Usage in Cryopreservation Process

Bhattacharya, Sankha

doi:10.5772/intechopen.80477

Cited by 59 publications

(80 citation statements)

References 28 publications

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“…Glycerol is a biocompatible monomer even at high concentrations, and it has little effect on enzymatic or metabolic processes [ 25 ]. As a result, glycerol is widely used as a cryoprotectant, where glycerol is dissolved in water and reduces freezing damage of various biomolecules [ 26 ]. Since glycerol consists of two primary and one secondary hydroxyl group, it serves as a trifunctional monomeric building for the synthesis of a wide range of hyperbranched polymers.…”

Section: Polyolsmentioning

confidence: 99%

Review on the Impact of Polyols on the Properties of Bio-Based Polyesters

et al. 2020

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Bio-based polyol polyesters are biodegradable elastomers having potential utility in soft tissue engineering. This class of polymers can serve a wide range of biomedical applications. Materials based on these polymers are inherently susceptible to degradation during the period of implantation. Factors that influence the physicochemical properties of polyol polyesters might be useful in achieving a balance between durability and biodegradability. The characterization of these polyol polyesters, together with recent comparative studies involving creative synthesis, mechanical testing, and degradation, have revealed many of their molecular-level differences. The impact of the polyol component on the properties of these bio-based polyesters and the optimal reaction conditions for their synthesis are only now beginning to be resolved. This review describes our current understanding of polyol polyester structural properties as well as a discussion of the more commonly used polyol monomers.

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Section: Polyolsmentioning

confidence: 99%

Review on the Impact of Polyols on the Properties of Bio-Based Polyesters

et al. 2020

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“…When replacing the water in a cell with a cryoprotector, the cell first shrinks due to osmosis, and then becomes swollen due to the penetration of the cryoprotectors into the cell. This is a classic reaction known in cryobiology as shrink swell [ 30 ]. The reaction proceeds until the volume of the cryoprotectors is equal to that of the original intracellular and extracellular fluids of the cells.…”

Section: Resultsmentioning

confidence: 99%

Long-Term Cryostorage of Mesenchymal Stem Cell-Containing Hybrid Hydrogel Scaffolds Based on Fibrin and Collagen

Egorikhina

Rubtsova

Aleynik

2020

Gels

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The most difficult issue when using tissue engineering products is enabling the ability to store them without losing their restorative capacity. The numbers and viability of mesenchymal stem cells encapsulated in a hydrogel scaffold after cryostorage at −80 °C (by using, individually, two kinds of cryoprotectors—Bambanker and 10% DMSO (Dimethyl sulfoxide) solution) for 3, 6, 9, and 12 months were determined, with subsequent assessment of cell proliferation after 96 h. The analysis of the cellular component was performed using fluorescence microscopy and the two fluorochromes—Hoechst 3334 and NucGreenTM Dead 488. The experimental protocol ensured the preservation of cells in the scaffold structure, retaining both high viability and proliferative activity during storage for 3 months. Longer storage of scaffolds led to their significant changes. Therefore, after 6 months, the proliferative activity of cells decreased. Cryostorage of scaffolds for 9 months led to a decrease in cells’ viability and proliferative activity. As a result of cryostorage of scaffolds for 12 months, a decrease in viability and proliferative activity of cells was observed, as well as pronounced changes in the structure of the hydrogel. The described scaffold cryostorage protocol could become the basis for the development of storage protocols for such tissue engineering products, and for helping to extend the possibilities of their clinical use while accelerating their commercialization.

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“…Among the most used are DMSO, DMF, MetOH glycerol and DMA, with a concentration that varies between 5 and 20% (usually 10%) of the volume [14], [24]. These chemicals, even if mildly toxic, have the property to surround the sperm cells, preventing the formation of ice crystals that could compromise membrane integrity and protecting them from cryodamage [25].…”

Section: Resultsmentioning

confidence: 99%

Optimization of cryopreservation andin vitrofertilization techniques for the African turquoise killifishNothobranchius furzeri

Dolfi

Suen

Ripa

et al. 2020

Preprint

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15Over the last decade, the African turquoise killifish, Nothobranchius furzeri, has 16 emerged as an important model system for the study of vertebrate biology and 17 ageing. However, rearing this fish in captivity can pose challenges, due to the 18 short window of fertility, inbreeding problems, and the continuous maintenance of 19 different strains and transgenic lines. To date, the main means of long term 20 strain maintenance is to arrest embryos in diapause, a poorly understood and 21 unreliable method. To solve these problems, we developed a robust protocol to 22 cryopreserve sperm and to revive them for in vitro fertilization (IVF), as a better 23 option for long term storage of N. furzeri lines. We tested a variety of extender 24 and activator buffers for sperm in vitro fertilization, as well as cryoprotectants to 25 achieve maximal long term storage and fertilization conditions tailored to this 26 species. Our optimized protocol was able to preserve sperm in a cryogenic 27 condition for months and to revive an average of 40% upon thawing. Thawed 28 sperm were able to fertilize nearly the same number of eggs as natural 29 fertilization, with an average of ~25% and peaks of ~55% fertilization. This 30 technical advance will greatly facilitate the use of N. furzeri as a model organism. 31 32 Abbreviations used: 33 HBSS: Hank's Balanced Salt Solution 34 BSMIS: Buffered sperm motility-inhibiting solution 35 FBS: Fetal bovine serum 3 36 DMSO: Dimethyl sulfoxide 37 DMF: Dimethylformamide 38 MetOH: Methanol 39 DMA: Dimethylacetamide 40 IVF: In vitro fertilization 41 42 Introduction 43 Over the last few years the African killifish, Nothobranchius furzeri, has emerged 44 as important model system for the study of vertebrate ageing. The life cycle of 45 these fish is characterized by a fast growth rate, reaching sexual maturity by 4-5 46 weeks, and a maximum lifespan of 6,5-7 months (as mean lifespan of the most 47 long-lived 10% of a given cohort)[1], [2], making them among the shortest-lived 48 vertebrate species bred in captivity and a unique platform for the rapid 49 exploration of aging and age-associated diseases[3]. 50 Unfortunately fast growth and aging features also carry some drawbacks that 51 make the maintenance in captivity of this species quite challenging. Rapid 52 growth and maturation lead to a rapid passing of successive generations, which 53 can often result in inbreeding [4], [5]. Indeed, it is virtually impossible to establish 54 an ancient founder to maintain an original genotype because of the marginal 55 overlap of generations. Fertility in this species is limited to the 5 th -20 th week of life 56 and is maximal between the 6 th and the 11 th [6]-[8], marking an extremely narrow 57 breeding window, especially compared to other fish species where this can 4 58 correspond to years. Moreover the success of transposon-mediated 59 transgenesis [9] and CRISPR-mediated mutagenesis [10] in this fish means that 60 more genetically engineered lines require continuous maintenance and space 61 u...

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Cryopretectants and Their Usage in Cryopreservation Process

Cited by 59 publications

References 28 publications

Review on the Impact of Polyols on the Properties of Bio-Based Polyesters

Review on the Impact of Polyols on the Properties of Bio-Based Polyesters

Long-Term Cryostorage of Mesenchymal Stem Cell-Containing Hybrid Hydrogel Scaffolds Based on Fibrin and Collagen

Optimization of cryopreservation andin vitrofertilization techniques for the African turquoise killifishNothobranchius furzeri

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