Cryopreservation of Agronomic Plant Germplasm Using Vitrification-Based Methods: An Overview of Selected Case Studies

Numerous environmental and endogenous factors affect the level of genetic diversity in natural populations. Genetic variability is the cornerstone of evolution and adaptation of species. However, currently, more and more plant species and local varieties (landraces) are on the brink of extinction due to anthropopression and climate change. Their preservation is imperative for the sake of future breeding programs. Gene banks have been created worldwide to conserve different plant species of cultural and economic importance. Many of them apply cryopreservation, a conservation method in which ultra-low temperatures (−135 °C to −196 °C) are used for long-term storage of tissue samples, with little risk of variation occurrence. Cells can be successfully cryopreserved in liquid nitrogen (LN) when the adverse effect of ice crystal formation and growth is mitigated by the removal of water and the formation of the so-called biological glass (vitrification). This state can be achieved in several ways. The involvement of key cold-regulated genes and proteins in the acquisition of cold tolerance in plant tissues may additionally improve the survival of LN-stored explants. The present review explains the importance of cryostorage in agronomy and presents an overview of the recent works accomplished with this strategy. The most widely used cryopreservation techniques, classic and modern cryoprotective agents, and some protocols applied in crops are considered to understand which parameters provide the establishment of high quality and broadly applicable cryopreservation. Attention is also focused on the issues of genetic integrity and functional genomics in plant cryobiology.

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Novel Approaches to the Cryopreservation of Human Spermatozoa: History and Development of the Spermatozoa Vitrification Technology

Journal of Reproductive and Stem Cell Biotechnology ◽

10.1177/205891581100200207 ◽

2011 ◽

Vol 2 (2) ◽

pp. 128-145 ◽

Cited By ~ 5

Author(s):

Evgenia Isachenko ◽

Gohar Rahimi ◽

Peter Mallmann ◽

Raul Sanchez ◽

Vladimir Isachenko

Keyword(s):

Ice Crystals ◽

Intracellular Lipids ◽

Cryoprotective Agents ◽

Long Term Storage ◽

Cryopreservation Techniques ◽

Water Quality And Quantity ◽

And Storage ◽

Term Storage ◽

Conventional Freezing

Cryobiology is very intensively applied in reproductive and veterinary medicine for preservation of gametes, embryos and reproductive tissues. Sub-zero temperatures combined with appropriate cryoprotective agents preserve the physiological and reproductive functions of the cells making long-term storage possible without loss of viability. With the use of cryoprotective agents it has become possible to develop cryopreservation techniques, such as the slow conventional freezing and vitrification that are in use in the present times. In slow controlled-rate conventional freezing extracellular ice crystals are formed whereas in vitrification no ice crystals are formed. Glass formation is compatible with the survival of the cell and the preservation of its intracellular structures provided the type(s) and concentrations of cryoprotectant used are not chemo- or osmotoxic. However, irrespective of the type of cooling method employed the cryosurvival of cells and tissues is influenced by the size and maturity of cells, amounts of intracellular water, quality and quantity of intracellular lipids, type of cells, their function and morphology. The intracellular milieu of cryopreserved cells and tissues remain less understood. The application of nanotechnology may help reveal and help advance our knowledge of the cryobiological principles involved in cryosurvival. At this moment the methods of cryopreservation that merit further investigation are vitrification and lyophilization. Vitrification is cheap if reagents are prepared in-house and the procedure can be performed rapidly. It has been successfully applied for gametes and embryos (of different stages of development), and reproductive cells/tissues, somatic cells and stem cells. However, vitrification is more demanding technically and requires operation and storage at sub-zero temperatures. On the other hand lyophilization deserves further investigation because it is a cheaper form of cryopreservation that may enable cryostorage at less demanding temperatures of 4°C and may even allow transport at ambient temperature. These possibilities are explored in this review.

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Fishes collected during the 2017 MarineGEO assessment of Kāne‘ohe Bay, O‘ahu, Hawai‘i

Journal of the Marine Biological Association of the United Kingdom ◽

10.1017/s0025315420000417 ◽

2020 ◽

Vol 100 (4) ◽

pp. 607-637

Author(s):

Lynne R. Parenti ◽

Diane E. Pitassy ◽

Zeehan Jaafar ◽

Kirill Vinnikov ◽

Niamh E. Redmond ◽

...

Keyword(s):

Smithsonian Institution ◽

Genetic Distances ◽

Taxonomic Identification ◽

Tissue Samples ◽

Long Term Storage ◽

Cytochrome Oxidase I Gene ◽

Mitochondrial Cytochrome Oxidase ◽

Term Storage ◽

I Gene

AbstractWe report the results of a survey of the fishes of Kāne‘ohe Bay, O‘ahu, conducted in 2017 as part of the Smithsonian Institution MarineGEO Hawaii bioassessment. We recorded 109 species in 43 families. The most speciose families were Acanthuridae (11 species), Gobiidae (11 species), Pomacentridae (10) and Chaetodontidae (9 species). Nine of the species that we collected are known or suspected to be introduced to the Hawaiian Islands. Specimens were identified, measured and photographed. All specimen vouchers were fixed in formalin and ultimately transferred to 75% ethanol for long-term storage. For nearly all species, we took multiple tissue samples from specimen vouchers prior to formalin-fixation; we preserved tissues in 95% ethanol and then stored them at −80°C. The 5′-end of the mitochondrial cytochrome oxidase I gene (mtCOI) was sequenced for 94 species to confirm their taxonomic identification. Using these barcode sequences, we also measured genetic distances between collected individuals and their conspecifics from other localities outside Hawaii to verify the hypothesis that Hawaiian populations of species broadly distributed throughout the Indo-Pacific may be genetically distinct. We present select case studies to demonstrate the potential for undiscovered endemism in the Hawaiian fish biota.

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Comparative analysis of the influence of cryoprotective agents and a long-term storage on human Adipose tissue-Mesenchimal Stromal Cells

Cytotherapy ◽

10.1016/j.jcyt.2020.03.176 ◽

2020 ◽

Vol 22 (5) ◽

pp. S100

Author(s):

O. Espinosa Ibáñez ◽

A. Fernández-González ◽

Á. Sierra-Sánchez ◽

J. Guerrero ◽

N. Fernández-Porcel ◽

...

Keyword(s):

Adipose Tissue ◽

Comparative Analysis ◽

Stromal Cells ◽

Human Adipose Tissue ◽

Cryoprotective Agents ◽

Long Term Storage ◽

Term Storage

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Suitability of Cryopreservation for the Long-term Storage of Rare and Endangered Plant Species: a Case History for Cosmos atrosanguineus

Annals of Botany ◽

10.1093/aob/mcg009 ◽

2002 ◽

Vol 91 (1) ◽

pp. 65-74 ◽

Cited By ~ 37

Author(s):

T. WILKINSON

Keyword(s):

Plant Species ◽

Case History ◽

Endangered Plant ◽

Endangered Plant Species ◽

Long Term Storage ◽

Rare And Endangered ◽

Term Storage

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Capture and Purification of Plant Genomic DNA on a Readily-available Cellulose Matrix

10.1101/163212 ◽

2017 ◽

Author(s):

Megan M. Thompson ◽

Estelle M. Hrabak

Keyword(s):

Arabidopsis Thaliana ◽

Nucleic Acids ◽

Plant Species ◽

Efficient Method ◽

Genomic Dna ◽

Low Cost ◽

Long Term Storage ◽

Cellulose Matrix ◽

Term Storage

AbstractWhatman FTA ®Cards are a fast and efficient method for capturing and storing nucleic acids but are cost-prohibitive for some researchers. We developed a method that substitutes readily-available cellulose-based paper and homemade washing buffer for commercial FTA ®Cards and FTA ®Purification Reagent. This method is suitable for long-term storage of DNA from many plant species, including Arabidopsis thaliana, prior to purification and PCR.Method SummaryHere we report a low-cost method for long-term storage of plant genomic DNA on a readily available cellulose matrix.

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Preservation of Avian Cells at Sub-zero Temperatures

Australian Journal of Biological Sciences ◽

10.1071/bi9790475 ◽

1979 ◽

Vol 32 (5) ◽

pp. 475 ◽

Cited By ~ 3

Author(s):

N Ratnamohan ◽

PB Spradbrow

Keyword(s):

Dimethyl Sulfoxide ◽

Cryoprotective Agent ◽

Cryoprotective Agents ◽

Long Term Storage ◽

Low Concentrations ◽

Short And Long Term ◽

Term Storage

The cryoprotective agents dimethyl sulfoxide (DMSO), glycerol, polyvinylpyrrolidone (PVP) and dextran were evaluated for their ability to protect avian cells during storage at sub-zero temperatures. DMSO was the most effective cryoprotective agent for the short- and long-term storage of avian cells and glycerol was also effective when used at low concentrations. PVP and dextran did not protect avian cells during storage in our experiments. Primary chicken cells and avian cells at higher passage levels were successfully recovered after storage with DMSO for periods ranging from 4 to 12 months.

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