Generation of Reactive Oxygen Species in Bovine Somatic Cell Nuclear Transfer Embryos during Micromanipulation Procedures

서지정보

· 발행기관 : 한국동물번식학회
· 수록지 정보 : Reproductive & developmental biology / 36권 / 1호
· 저자명 : In-Sun Hwang, Hyo-Kyung Bae, Choon-Keun Park, Boo-Keun Yang, Hee-Tae Cheong

목차

ABSTRACT INTRODUCTION MATERIALS AND METHODS In Vitro Maturation of Oocytes Culture of Somatic Cells Nuclear Transfer Electrofusion and Activation In Vitro Fertilization (IVF) In Vitro Culture of Embryos Analysis of ROS Products Statistical Analysis RESULTS ROS Generation Levels during Manipulation Procedures ROS Generation Levels during Early In Vitro Culture of SCNT and IVF Embryos DISCUSSIONS REFERENCES

영어초록

The present study was conducted to examine the generation of reactive oxygen species (ROS) during micromanipulation procedures in bovine somatic cell nuclear transfer (SCNT) embryos. Bovine enucleated oocytes were electrofused with donor cells, activated by a combination of Ca-ionophore and 6-dimethylaminopurine culture. Oocytes and embryos were stained in dichlorodihydrofluorescein diacetate or 3'-(p-hydroxyphenyl) fluorescein dye and the H2O2 or ˙OH radical levels were measured. In vitro fertilization (IVF) was performed for controls. The samples were examined with a fluorescent microscope, and fluorescence intensity was analyzed in each oocyte and embryo. The H2O2 and ˙OH radical levels of reconstituted oocytes were increased during manipulation (37.2~49.7 and 51.0~55.2 pixels, respectively) as compared to those of mature oocytes (p<0.05). During early in vitro culture, the ROS levels of SCNT embryos were significantly higher than those of IVF embryos (p<0.05). These results suggest that the cellular stress during micromanipulation procedures can generate the ROS in bovine SCNT embryos.

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