In vitro development of bovine one-cell embryos fertilized in vitro in serum- and feeder cell-free culture systems |
S. Ohboshi, K. Hanada, J. Zhao, M. Hattori, N. Fujihara, R. Umetsu, T. Yoshida, H. Tomogane |
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Abstract |
The purpose of this study was to evaluate some factors in the bovine embryonic development from one-cell to blastocyst using modified synthetic oviduct fluid medium(mSOFM), after maturation and in vitro fertilization of the oocytes. The embryonic development to the blastocyst stage was assessed at 7-10 days after in vitro fertilization, and the total cells in the blastocysts were counted by staining nuclei with fluorochrome. Some commercial calf sera(CS) and a superovulated cow serum had different effects on the embryonic development to the blastocyst stage(8.6-21.4%), dependent ipon their product lots, although the development might not be affected at least by serum progesterone levels. 棺-Mercaptoethanol(棺-ME) supplemented into mSOFM was effective to the embryonic development(27.8%), as well as the co-culture system with cumulus cells(19.5%). In a serum- and feeder cell-free culture using mSOFM containing several growth factors and 棺-ME instead of CS plus co-cultured cumulus cells, bovine serum albumin(BSA, fraction V), but not polyvinyl alcohol(PVA), was highly effective in embryonic development to the blastocyst stage, almost comparable to CS in the serum-contained culture(CS, BSA and PVA; 27.8, 19.5 and 5.7%, respectively). However, fatty acid free BSA rather reduced the number of developed blastocysts, compared with fraction V BSA(7.3 vs 29.4%). In the serum- and feeder cell-free culture, supplement of glucose to the medium(final 2.0mM) stimulated the cell proliferation of developing embryos 120 hr after in vitro fertilization. These results indicated that a serum-free medium supplemented with 棺-ME could successfully support the development of bovine one-cell embryos to the blastocyst stage. Moreover, supplement of glucose and fatty acids to the medium might support preferably the development and cell proliferation of embryos. |
Keywords:
Bovine Embryonic Development; Serum-free Medium; 棺-Mercaptoethanol; Bovine Serum Albumin |
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