원문정보
초록
영어
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.
목차
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