Cryo-mesh: A simple alternative technique for cryopreservation

Funnekotter, Bryn; Bunn, Eric; Mancera, Ricardo L.

doi:10.1016/j.cryobiol.2016.09.163

Cited by 11 publications

(21 citation statements)

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“…(b) Multiple shoot tips on each cryo-mesh can then be exposed to cryoprotective agents. (c) Cryo-mesh plunged into liquid nitrogen for cryo-storage (Funnekotter et al 2017a).…”

Section: Discussionmentioning

confidence: 99%

“…The vitrification protocol is currently the most commonly used cryopreservation technique, and hundreds of plant species have been successfully cryopreserved in this way (Sakai and Engelmann 2007). The vitrification method has continued to develop and improve over time, with approaches such as vacuum infiltration vitrification (VIV) (Nadarajan and Pritchard 2014;Funnekotter et al 2015), cryo-plate protocols (Yamamoto et al 2011(Yamamoto et al , 2012, and a new cryo-mesh (Funnekotter et al 2017a) protocol created to increase success in cryopreservation.…”

Section: Advancements In Cryopreservation Protocolsmentioning

confidence: 99%

“…Yamamoto et al (2012) achieved post-cryopreservation regrowth between 73-100% across 16 species. Funnekotter et al (2017a) investigated an alternative to the cryo-plate protocol in which an aluminium mesh, deemed cryo-mesh, was used in place of the cryo-plate with fixed well sizes. Cryo-plates require specific and consistent production of wells or shoot tips may not be able to adhere.…”

Section: Advancements In Cryopreservation Protocolsmentioning

confidence: 99%

“…Cryo-mesh overcomes this by allowing the alginate to act as a glue between the mesh and the shoot tips ( Fig. 5) (Funnekotter et al 2017a). Cryo-mesh efficacy was tested in comparison to droplet-vitrification with cryopreservation of the West Australian native species A. viridis.…”

Section: Advancements In Cryopreservation Protocolsmentioning

confidence: 99%

“…No significant difference in post-cryogenic generation was observed between the two protocols: 78% for droplet-vitrification and 83% for cryo-mesh. Cryo-mesh, however, showed efficacy in reducing the manufacturing precision required over cryo-plates, as well as reducing the operator skill required in comparison to the dropletvitrification protocol (Funnekotter et al 2017a). Meristem shoot tips from tissue cultured plants are commonly used in cryopreservation and are desiccated by osmotic means; however, excised zygotic embryos and embryonic axes are the preferred materials for cryopreservation as they allow for whole plants to be generated without complex tissue culture protocols (Ballesteros et al 2014).…”

Section: Advancements In Cryopreservation Protocolsmentioning

confidence: 99%

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Current issues in plant cryopreservation and importance for ex situ conservation of threatened Australian native species

et al. 2019

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An alarming proportion of Australia’s unique plant biodiversity is under siege from a variety of environmental threats. Options for in situ conservation are becoming increasingly compromised as encroaching land use, climate change and introduced diseases are highly likely to erode sanctuaries regardless of best intentions. Ex situ conservation is currently limited to botanic garden living collections and seed banking, with in vitro and cryopreservation technologies still being developed to address ex situ conservation of species not amenable to conventional storage. Cryopreservation (storage in liquid nitrogen) has been used successfully for long-term biosecure storage of shoot tips of several species of threatened Australian plants. We present a case for building on this research and fostering further development and utilisation of cryopreservation as the best means of capturing critical germplasm collections of Australian species with special storage requirements (e.g. recalcitrant-seeded taxa and species with short-lived seeds) that currently cannot be preserved effectively by other means. This review highlights the major issues in cryopreservation that can limit survival including ice crystal damage and desiccation, toxicity of cryoprotective agents, membrane damage, oxidative stress and mitochondrial function. Progress in understanding and mitigating these stresses is vital for advancing cryopreservation for conservation purposes.

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“…(b) Multiple shoot tips on each cryo-mesh can then be exposed to cryoprotective agents. (c) Cryo-mesh plunged into liquid nitrogen for cryo-storage (Funnekotter et al 2017a).…”

Section: Discussionmentioning

confidence: 99%

Section: Advancements In Cryopreservation Protocolsmentioning

confidence: 99%

Section: Advancements In Cryopreservation Protocolsmentioning

confidence: 99%

Section: Advancements In Cryopreservation Protocolsmentioning

confidence: 99%

Section: Advancements In Cryopreservation Protocolsmentioning

confidence: 99%

See 3 more Smart Citations

Current issues in plant cryopreservation and importance for ex situ conservation of threatened Australian native species

et al. 2019

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Conduction‐Dominated Cryomesh for Organism Vitrification

Guo,

Zuchowicz,

Bouwmeester

et al. 2023

Advanced Science

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Vitrification‐based cryopreservation is a promising approach to achieving long‐term storage of biological systems for maintaining biodiversity, healthcare, and sustainable food production. Using the “cryomesh” system achieves rapid cooling and rewarming of biomaterials, but further improvement in cooling rates is needed to increase biosystem viability and the ability to cryopreserve new biosystems. Improved cooling rates and viability are possible by enabling conductive cooling through cryomesh. Conduction‐dominated cryomesh improves cooling rates from twofold to tenfold (i.e., 0.24 to 1.2 × 105 °C min−1) in a variety of biosystems. Higher thermal conductivity, smaller mesh wire diameter and pore size, and minimizing the nitrogen vapor barrier (e.g., vertical plunging in liquid nitrogen) are key parameters to achieving improved vitrification. Conduction‐dominated cryomesh successfully vitrifies coral larvae, Drosophila embryos, and zebrafish embryos with improved outcomes. Not only a theoretical foundation for improved vitrification in µm to mm biosystems but also the capability to scale up for biorepositories and/or agricultural, aquaculture, or scientific use are demonstrated.

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Exogenous preculture with sucrose and abscisic acid improves post-cryopreservation survival of eastern bracken fern gametophytes

Jang,

Oh,

Kim

et al. 2023

Sci Rep

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Cryopreservation is an important technique used in the conservation of various plant tissues. This study proposes a cryopreservation method for the long-term conservation of eastern bracken fern gametophytes (Pteridium aquilinum var. latiusculum). Encapsulation–dehydration of the gametophytes was performed, and the exogenous sucrose and abscisic acid (ABA) preculture conditions were investigated. Gametophytes are sensitive to dehydration and drying, and the following treatment conditions were applied: encapsulation by alginate containing 0.75 M sucrose, 18-h loading treatment with 0.75 M sucrose, and 6-h drying treatment. The survival rate following cryopreservation was determined. The water content of < 27.5% in the alginate beads after dehydration and drying was found to be appropriate for ensuring survival. Additionally, performing an exogenous sucrose and ABA preculture was essential before encapsulation to achieve a survival of ≥ 90%. The high stress induced by cryopreservation and exogenous preculture regulated the expression of PaSuSy, PaLEA14, and PaABI1b and the endogenous ABA content. In eastern bracken gametophytes, ABI1 appears to be a negative regulator of ABA signaling. These results indicate that the encapsulation–dehydration method is effective for the long-term conservation of eastern bracken fern gametophytes, and exogenous preculture alleviates abiotic stress and increases the survival rate.

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Cryo-mesh: A simple alternative technique for cryopreservation

Cited by 11 publications

References 0 publications

Current issues in plant cryopreservation and importance for ex situ conservation of threatened Australian native species

Current issues in plant cryopreservation and importance for ex situ conservation of threatened Australian native species

Conduction‐Dominated Cryomesh for Organism Vitrification

Exogenous preculture with sucrose and abscisic acid improves post-cryopreservation survival of eastern bracken fern gametophytes

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