TITLE

The effects of vitrification on nuclear status of ovine oocytes

AUTHOR(S)
Taheri, F.; Shirazi, A.; Nazari, H.; Ahmadi, E.; Mangoli, E.
PUB. DATE
June 2014
SOURCE
Iranian Journal of Reproductive Medicine;Jun2014 Supplement, Vol. 12, p74
SOURCE TYPE
Academic Journal
DOC. TYPE
Abstract
ABSTRACT
Introduction: The microtubules and chromatin of oocytes are vulnerable to cryoprotectants and thermal change during cryopreservation. The objective of the present study was to investigate the effects of vitrification on nuclear status of ovine oocytes in GV and Mil developmental stages. Materials and Methods: The oocytes from follicles of ovine, 2-6 mm in diameter, were collected in a pre-incubated (38.6°C, 5% CO2, 100% humidity) HEPES-modified TCM 199 medium. Good quality oocytes were either vitrified in GV or after IVM as Mil stage. The basic vitrification solution was HEPES-buffered TCM-199 with FCS. The selected oocytes were placed in equilibration solution of 7.5% EG + 7.5% DMSO for 5 min, then transferred into vitrification solution containing 15% EG, 15% DMSO, and 0.5M sucrose for 30 sec. The oocytes were then loaded onto the tip of modified straw and plunged into liquid nitrogen. Following warming in solution containing 0.5 M sucrose, vitrified GV and Mil oocyte were cultured in IVM medium for 24h and 2h, respectively. Then, they were denuded, and stained with Hoechst. They were then evaluated for the chromatin status under a fluorescence microscope. Results: The rate of Mil oocyte with normal nuclear status in vitrified-warmed GV and Mil oocyte were no statistically significant (43% and 41%, respectively; p<0.05) but the rate of Mil oocyte with abnormal nuclear status in vitrified-warmed Mil oocytes (55%) was significantly (p<0.05) higher than those of vitrified-warmed GV oocytes (41%). Conclusion: The chromatin of Mil oocytes was more sensitive to vitrification than those of GV oocytes.
ACCESSION #
96841663

 

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