원문정보
초록
영어
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.
목차
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
REFERENCES