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Cryobiology
Article . 2011 . Peer-reviewed
License: Elsevier TDM
Data sources: Crossref
Cryobiology
Article . 2011
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Advances in the cryopreservation of sea-urchin embryos: Potential application in marine water quality assessment

Authors: Bellas, J. (Juan); Paredes, E. (Estefanía);

Advances in the cryopreservation of sea-urchin embryos: Potential application in marine water quality assessment

Abstract

Among the most widely used biological techniques in marine pollution assessment, the sea-urchin embryo-larval bioassay is in an advanced developmental stage. Cryopreservation might help to overcome the problem of the spawning seasonality and therefore strengthen the use of those embryo-larval bioassays. This work investigates different factors influencing cryopreservation of sea-urchin embryos, including the cooling rates and holding temperatures, the seeding, or the impact of plunging into liquid nitrogen. The blastula stage yielded better results than the fertilised egg, and the most effective cryoprotectants combination was dimethyl sulfoxide 1.5M plus trehalose 0.04M. The optimised protocol developed begins with an initial hold at 4°C for 2min, followed by cooling at -1°Cmin(-1) to -12°C. At this point a seeding step was incorporated with a 2min hold, followed by a second cooling at -1°Cmin(-1) to -80°C. After a final hold of 2min the cryovials are transferred into liquid nitrogen for storage. Although the cryopreservation processes might cause a delay in the development of sea-urchin embryos, high larval growth (71-98% of controls) was obtained when cryopreserved blastulae were further incubated for 72-96h in artificial seawater. We conclude that embryo-larval bioassays with cryopreserved sea-urchin blastulae are suitable for use in marine pollution monitoring programs and may be considered as an acceptable solution to the reproductive seasonality of sea-urchin species.

Country
Spain
Keywords

Male, Embryo, Nonmammalian, Time Factors, Cryoprotective Agents, Centro Oceanográfico de Vigo, Animals, Dimethyl Sulfoxide, Seawater, Medio Marino, Cryopreservation, Water Pollution, Temperature, Trehalose, Blastula, Larva, Sea Urchins, Paracentrotus, Biological Assay, Female, Environmental Monitoring

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visibility
selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
views
OpenAIRE UsageCountsViews provided by UsageCounts
35
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