PUBLICATION
Cryopreservation of zebrafish (Danio rerio) oocytes by vitrification
- Authors
- Guan, M., Rawson, D.M., and Zhang, T.
- ID
- ZDB-PUB-101011-43
- Date
- 2010
- Source
- Cryo letters 31(3): 230-238 (Journal)
- Registered Authors
- Rawson, David M., Zhang, Tiantian
- Keywords
- zebrafish, vitrification, trypan blue staining
- MeSH Terms
-
- Animals
- Cryopreservation/methods*
- Cryoprotective Agents*
- Oocytes*
- Tissue Survival
- Zebrafish*
- PubMed
- 20919452
Citation
Guan, M., Rawson, D.M., and Zhang, T. (2010) Cryopreservation of zebrafish (Danio rerio) oocytes by vitrification. Cryo letters. 31(3):230-238.
Abstract
Cryopreservation of fish oocytes is challenging because these oocytes have low membrane permeability to water and cryoprotectant and are highly chilling sensitive. Vitrification is considered to be a promising approach for their cryopreservation as it involves rapid freezing and thawing of the oocytes and therefore minimising the chilling injury. In the present study, vitrification properties and the toxicity of a range of vitrification solutions containing different concentrations of Me2SO, methanol, propylene glycol and ethylene glycol were investigated. Two different base media and vitrification methods were compared. The effect of different post-thaw dilution solutions together with incubation periods on oocyte viability were also investigated. Stage III zebrafish oocytes were equilibrated in increasing concentrations of cryoprotectants for 30 min in 3 steps. Oocytes were thawed rapidly in a water bath and cryoprotectants were removed in 4 steps. Oocyte viability was assessed using trypan blue staining. The results showed that vitrification solutions V3 and V4 in KCl buffer had low toxicity and vitrified well. The survivals of oocytes after stepwise dilution using solutions containing permeable cryoprotectants were significant higher than those diluted in 0.5M glucose, and the use of CVA65 vitrification system improved oocyte survival when compared with plastic straws after 30 min at 22 degrees C post-thawing. Cryopreservation of zebrafish oocytes by vitrification is reported here for the first time, although oocyte survivals after cryopreservation assessed by trypan blue staining were relatively high shortly after thawing, they became swollen and translucent after incubation in KCl buffer. Further studies are needed to optimise the post-thaw culturing conditions.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping