Cryopreservation of zebrafish (Danio rerio) blastomeres using controlled slow cooling

Abstract

Cryopreservation of aquatic species has been widely studied especially zebrafish gametes, embryos and larvae. Cryopreservation of blastomeres has the advantage of preserving both paternal and maternal genetic information. The research work presented in this thesis investigated the toxicity of cryoprotectants to 75% epiboly stage blastomeres of zebrafish (Danio rerio) before cryopreservation. DMSO was found to be the least toxic cryoprotectant for blastomeres, after 30 min incubated in PBS at room temperature, with 97.8% survivaL Cryopreservation of zebrafish blastomeres was carried out using controlled slow cooling. In addition to cryoprotectant, the cryoprotective properties of other compounds were also investigated including NaHC03, coffee and honey, and their solutions were used as cryopreservation media. Comparison of blastomere survival in different media after freezing showed honey to be the most effective, with 98.1 % survival immediately after freeze thawing. Comparison of blastomers cryopreserved in honey and DMSO, after 60 min incubation in PBS at room temperature following freeze-thawing and cryoprotectant removal, showed honey to be the more effective cryoprotectant for controlled slow cooling of zebrafish blastomeres with 97.2% survival. Results from the 'present study showed honey to have properties that protect the blastomere from freezing injury.

Date of Award	Jul 2008
Original language	English
Awarding Institution	University of Bedfordshire