Effect of Porcine Follicular Fluid on Donor Cell Characteristics and Quality of Porcine Cloned Blastocysts
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서지정보
ㆍ발행기관 : 한국동물번식학회
ㆍ수록지정보 : Reproductive & developmental biology / 36권 / 4호
ㆍ저자명 : Dae-Jin Kwon, Keon Bong Oh, Sun A Ock, Sung-Soo Lee, Jin-Ki Park, Won-Kyong Chang, Seongsoo Hwang, Jeong Woong Lee
ㆍ저자명 : Dae-Jin Kwon, Keon Bong Oh, Sun A Ock, Sung-Soo Lee, Jin-Ki Park, Won-Kyong Chang, Seongsoo Hwang, Jeong Woong Lee
목차
ABSTRACT INTRODUCTION MATERIALS AND METHODS Preparation of Cumulus-Oocyte Complexes (COCs) and FF Isolation and Culture of Miniature Pig Ear Fibroblasts Determination of Amino Acid Concentrations Cell Proliferation Assay Determination of Apoptotic and Necrotic Cell Distribution SCNT Procedure TUNEL Assay for Apoptotic Cell Death Statistical Analysis RESULTS Measurement of Amino Acid Concentration Cell Proliferation Assay Determination of Apoptotic and Necrotic Cells Developmental Competence of Cloned Embryos Apoptotic Cell Death in Cloned Blastocysts DISCUSSION ACKNOWLEDGEMENTS REFERENCES영어 초록
This study aimed at investigating whether a porcine follicular fluid (pFF) supplementation positively affects the characteristics of donor cells and the developmental competence of porcine cloned embryos. Ear fibroblast cells (donor cell) from an Massachusetts General Hospital miniature pig were cultured in different culture methods: (1) Dulbecco's modified Eagle's medium (DMEM)+10% FBS (Control); (2) DMEM+0.5% FBS (SS); and (3) DMEM+10% FBS+10% pFF (pFF) for 72 h. In each conditioned medium, the concentrations of 4 amino acids (Thr, Glu, Pro, and Val) in the pFF group were significantly different from those in the control group (p<0.05 or p<0.01). The proliferation of the cells cultured in the SS group was significantly lower than that of the other treatment groups (p<0.01). The population of apoptotic and necrotic cells in the SS group was significantly higher than that of either the control or the pFF group (p<0.01). The number of embryos that cleaved (p<0.05) and developed into blastocysts (p<0.01) in the SS group was significantly lower than that of either the control or the pFF group. Compared to other groups, the blastocysts produced from the donor cells in the pFF group had higher total cells and lower apoptotic cells (p<0.05). It can be concluded that pFF supplementation in the donor cell culture medium positively affects cell death, cell cycle and quality of the cloned blastocyst.참고 자료
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