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    Short-term chilled storage of Zebrafish (Danio rerio) embryos in cryoprotectant as an alternative to cryopreservation

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    Authors
    Desai, Kunjan
    Spikings, Emma
    Zhang, Tiantian
    Affiliation
    Georgia Regents University
    University of Bedfordshire
    Bournemouth University
    Issue Date
    2015-02
    Subjects
    zebrafish
    Danio rerio
    cryopreservation
    
    Metadata
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    Abstract
    As zebrafish embryos have never been cryopreserved, we developed a protocol to store zebrafish embryos (50% epiboly-5.3 hour post fertilization) for up to 18 h at 0°C. Initial experiments to optimize the cryoprotectant (CPA) solution demonstrated improved embryo hatching rate following chilling at 0°C for 18 h with 1 M MeOH+0.1 M sucrose (56 ± 5%) compared with other combinations of methanol (0.2-0.5 M) and sucrose (0.05-0.1 M). This combination of CPAs that protects against chilling injury was further tested to assess its impact on sox gene and protein expression. Significant decreases in sox3 gene expression were observed in hatched embryos that had been chilled for 18 h in 1 M MeOH+0.1 sucrose compared with non-chilled controls, however the expression of both sox2 and sox3 proteins was unaffected. Significant decreases in sox2 protein expression were, however, observed in embryos that had been chilled without CPAs and these embryos also had lower hatching rates than those chilled with the optimal CPA solution. We, therefore, conclude that the CPA combination of 1 M MeOH+0.1 M sucrose facilitates chilled storage of early stage (50% epiboly) zebrafish embryos for up to 18 h without compromising transcriptional response.
    Citation
    Desai, K., Spikings, E. and Zhang, T.T. (2015) 'Short-term chilled storage of Zebrafish (Danio rerio) embryos in cryoprotectant as an alternative to cryopreservation' Zebrafish 12(1), pp.111-120.
    Publisher
    Mary Ann Liebert
    Journal
    Zebrafish
    URI
    http://hdl.handle.net/10547/560929
    DOI
    10.1089/zeb.2013.0961
    Additional Links
    http://online.liebertpub.com/doi/abs/10.1089/zeb.2013.0961
    Type
    Article
    Language
    en
    ISSN
    1545-8547
    1557-8542
    ae974a485f413a2113503eed53cd6c53
    10.1089/zeb.2013.0961
    Scopus Count
    Collections
    Biomedical Science

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