Publication: Developmental rates of vitrified buffalo oocytes following parthenogenetic activation and intracytoplasmic sperm injection
Issued Date
2010-01-01
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ISSN
16696840
16684834
16684834
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2-s2.0-84904721040
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Mahidol University
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SCOPUS
Bibliographic Citation
Revista Veterinaria. Vol.21, No.SUPPL.1 (2010), 856-863
Suggested Citation
Yuanyuan Liang, Tatsanee Phermthai, Takashi Nagai, Tamas Somfai, Rangsun Parnpai Developmental rates of vitrified buffalo oocytes following parthenogenetic activation and intracytoplasmic sperm injection. Revista Veterinaria. Vol.21, No.SUPPL.1 (2010), 856-863. Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/28552
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Title
Developmental rates of vitrified buffalo oocytes following parthenogenetic activation and intracytoplasmic sperm injection
Abstract
The objective of this study was to investigate the potential of swamp buffalo oocytes vitrified-warmed at the MII stage to develop to the blastocyst stage after parthenogenetic activation (PA) and intracytoplasmic sperm injection (ICSI). In the first experiment, we examined the toxic effects of cryoprotectants on in vitro development. In vitro matured oocytes were placed in 10% dimethylsulfoxide (DMSO) + 10% ethylene glycol (EG) for 1 min and then exposed to 20% DMSO + 20% EG + 0.5 M sucrose for 30 sec (1+30), 45 sec (1+45) or 60 sec (1+60). The oocytes were exposed to warming solution (0.5M sucrose) for 5 min and then washed in TCM199 HEPES + 20%FBS for 5 min. Oocyte viability was assessed by fluorescein diacetate (FDA) staining. Surviving oocytes were parthenogenetically activated and cultured for 7 days. The viability in all groups of CPA treatment and the control were 100%. The development rates to the blastocyst stage among CPA exposed 1+30 (17%), control (23%) and fresh control (control without FDA assay) (27%) did not differ significantly, but they were significantly higher than those in CPA exposed 1+45 (9%) and 1+60 (1%) groups. In the second experiment, we examined the effect of two CPA exposure times, 1+30 and 1+45 on the in vitro development for 7 days after PA of oocytes vitrified by the microdrop method. The viability in vitrified 1+30, 1+45 and the control groups was not different (97%, 95% and 100%, respectively). The development of surviving oocytes to blastocyst stage in the vitrified 1+30 group (8%) was significantly higher than that in the vitrified 1+45 group (4%) and significantly lower than those in control and fresh control groups (24% and 26%, respectively). In the third experiment, we examined the effect of two CPA exposure times, 1+30 and 1+45 on in vitro development after ICSI of vitrified oocytes. The FDA viability in vitrified 1+30, 1+45 and control groups (100%) was not different (96%, 91% and 100%, respectively). After ICSI vitrified-warmed oocytes were activated and oocytes with the 2nd polar body were cultured for 7 days. The development of ICSI oocytes to the blastocyst stage in the vitrified 1+30 group (11%) was significantly higher than that in vitrified 1+45 (7%) and significantly lower than those in the control and fresh control (21% and 23%, respectively). In conclusion, our study demonstrated that the 1+30 CPA treatment regimen could yield the highest blastocyst rates for oocytes vitrified by the microdrop method and that the FDA viability test had no effect on the embryo development.