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Effect of Porcine Follicular Fluid on Donor Cell Characteristics and Quality of Porcine Cloned Blastocysts

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한국동물번식학회 (The Korean Society of Animal Reproduction)
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.

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   REFERENCES
저자
  • Dae-Jin Kwon(Animal Biotechnology Division, National Institute of Animal Science, RDA, Suwon 441-706, Korea)
  • Keon Bong Oh(Animal Biotechnology Division, National Institute of Animal Science, RDA, Suwon 441-706, Korea)
  • Sun A Ock(Animal Biotechnology Division, National Institute of Animal Science, RDA, Suwon 441-706, Korea)
  • Sung-Soo Lee(Animal Biotechnology Division, National Institute of Animal Science, RDA, Suwon 441-706, Korea)
  • Jin-Ki Park(Animal Biotechnology Division, National Institute of Animal Science, RDA, Suwon 441-706, Korea)
  • Won-Kyong Chang(Animal Biotechnology Division, National Institute of Animal Science, RDA, Suwon 441-706, Korea)
  • Seongsoo Hwang(Animal Biotechnology Division, National Institute of Animal Science, RDA, Suwon 441-706, Korea) Corresponding author
  • Jeong Woong Lee(Regenerative Medicine Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-806, Korea)