Cryoloop vitrification of rabbit oocytes

BACKGROUND: Vitrification is assumed to be a promising method to cryopreserve human oocytes but still needs optimization. In this study, rabbit oocytes (fertilized by ICSI) were vitrified with cryoloops, and the effect of three different cryopreservation protocols on spindle configuration and embryo...

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Bibliographic Details
Published in:Human reproduction (Oxford) 2005-07, Vol.20 (7), p.1969-1974
Main Authors: Cai, X.Y., Chen, G.A., Lian, Y., Zheng, X.Y., Peng, H.M.
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Chromosomes - ultrastructure
cryoloop
cryopreservation
Cryopreservation - methods
Cryoprotective Agents
Dimethyl Sulfoxide
Embryonic Development
Ethylene Glycol
Female
Gynecology. Andrology. Obstetrics
Humans
In Vitro Techniques
Medical sciences
Metaphase
Microtubules - ultrastructure
oocyte/rabbit
Oocytes - cytology
Rabbits
Sperm Injections, Intracytoplasmic
spindle
Spindle Apparatus - ultrastructure
vitrification
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Summary:BACKGROUND: Vitrification is assumed to be a promising method to cryopreserve human oocytes but still needs optimization. In this study, rabbit oocytes (fertilized by ICSI) were vitrified with cryoloops, and the effect of three different cryopreservation protocols on spindle configuration and embryo quality was assessed. METHODS: Metaphase II rabbit oocytes were randomly assigned to one of four groups: (i) control; (ii) E40 [40% ethylene glycol (EG)]; (iii) ED20 [20% EG + 20% dimethylsulphoxide (DMSO)]; and (iv) ED20 + M (20% EG+20% DMSO+ vitrification machine). After warming, one part of each group was fertilized by ICSI to examine the fertilization and embryo cleavage ability, and the others were immunostained for tubulin and chromatin before visualization using confocal microscopy. RESULTS: The survival rates after warming were 79.1, 83.1 and 82.3%, respectively. In protocols E40 and ED20, the spindles were severely injured and the embryo quality not good compared with those in the ED20 + M group. CONCLUSIONS: The fastest cooling rate in combination with EG and DMSO as cryoprotectants had the fewest adverse effects on the spindle configuration of rabbit oocytes and embryo development.
ISSN:0268-1161
1460-2350
DOI:10.1093/humrep/deh805