Study on Development of Canine Oocytes Treated by In Vitro Fertilization and ICSI
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서지정보
ㆍ발행기관 : 한국동물번식학회
ㆍ수록지정보 : Reproductive & developmental biology / 35권 / 2호
ㆍ저자명 : Ji-Hoon Park, Young-Ho Chung, Sang Keun Kim
ㆍ저자명 : Ji-Hoon Park, Young-Ho Chung, Sang Keun Kim
목차
ABSTRACT INTRODUCTION MATERIALS AND METHODS Recovery and Culture of Oocytes Vitrification and Thawing of Oocytes IVF Procedure ICSI Procedure Assessment of Survival and Developmental Rate Statistical Analysis RESULTS Viability of Fresh and Vitrified-Thawed Oocytes IVM and IVF of Fresh and Vitrified Oocytes In Vitro Development of Oocytes Following IVF and ICSI DISCUSSION REFERENCES영어 초록
These study was to investigate the in vitro fertilization and viability of fresh and vitrified oocytes. Also, the developmental capacity of IVF and intracytoplasmic sperm injection (ICSI) oocytes were investigated. Then vitrification was performed with the use of 20% ethylene glycol + 20% DMSO + 0.5 M sucrose + 10% FCS + TCM-199 medium. Vitrification immature oocytes are cultured in vitrification solution for 10 min afterwards transferred to expose at room temperature for 5 min. and transferred to the ice water for 5 min. The oocytes were sealed in a 1.0 mm straw and placed in a LN2 container. Frozen oocytes were rapidly thawed in a water bath at 30~35℃, and then placed in TCM-199 medium containing 0.5 M sucrose for 5 min each, respectively, at 38℃. After being washed for 2~3 times, using fresh medium the oocytes were cultured in TCM-199 medium supplemented with 5% FCS at 38℃ in 5% CO2 and air. The normal morphology of fresh and vitrified-thawed oocytes were 87.1±2.1% and 54.8±2.5%, respectively. The viability rates of fresh and vitrified-thawed oocytes were 70.0±2.2% and 41.9±2.6%, respectively. Viability rates of vitrified-thawed oocytes were lower than that of fresh follicular oocytes (p<0.05). The in vitro maturation rates of fresh and vitrified oocytes were 45.1±3.6% and 28.9±4.4%, respectively. The IVF rates of fresh follicular and vitrified-thawed oocytes were 34.0±2.2% and 20.2±2.6%, respectively. The in vitro maturation and fertilization rates of vitrified-thawed oocytes were lower than those of the fresh follicular oocytes (p<0.05). A total of 350 oocytes were fixed and stained after co-incubation with spermatozoa, of which 88 had identifiable nuclear material. After IVF for 20 hrs, 25.1±3.4% of the oocytes found to have been penetrated by spermatozoas. Oocytes were fixed and stained after ICSI, and 105 oocytes contained identifiable nuclear material. After IVF and ICSI for 20 hrs, 34.3±3.4% and 59.0±2.0% of the oocytes were found to have been penetrated by spermatozoas. The developmental rates upon ICSI were significantly higher than those of the IVF method (p<0.05).참고 자료
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