The storage of culture media (M16) by freezing or freeze-drying for the development of mouse embryos
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概要
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The present study was undertaken to examine the effects of culture media (M16) stored by freezing (-20°C) or freeze-drying on the development of mouse zygotes or 2-cell embryos in vitro and in vivo.<BR>Zygotes and 2-cell embryos were obtained from the superovulated CD-1 female mice mated with CD-1 males on 19-23 or 43-46 hrs after hCG injection, respectively. Zygotes with two pronuclei were cultured for 4 days with fresh and frozen media supplemented with 100 μM EDTA. Two-cell embryos were cultured for 3 days with fresh, frozen and freeze-drying media. Some blastocysts developed from zygotes or 2-cell embryos cultured with each media were transferred into the oviducts on day 0.5 or into the uteri on day 2.5 or 3.5 of pseudopregnant mice. They were killed on day 16.5 or 17.5 to examine the number of live fetuses. Cell numbers in both trophectoderm and inner cell mass of blastocysts on freeze-drying media were also examined.<BR>High proportions of zygotes and 2-cell embryos cultured with frozen and freeze-drying media developed to blastocysts in vitro (59% and 86% for frozen medium, 89% for freeze-drying medium), and were not significantly differentcompared with those obtained in fresh medium (59% and 86%).<BR>The proportion of live fetuses after transfer of blastocysts developed from zygotes or 2-cell embryos with frozen medium (34% and 31%) were not significantly different from those obtained with fresh medium (35% and 39%, respectively). However, only 8% of embryos cultured with freeze-drying medium developed to fetuses. But, the cell numbers in trophectoderm and inner cell mass of blastocysts were not significantly different from those obtained with fresh medium.<BR>The present study demonstrated that M16 medium could be preserved at least for 10 months at -20°C without the decrease of its potency for the development of mouse embryos in vitro and in vivo. However, although the reason was not clear, freeze-drying of medium resulted in a drastical decrease in the percentage of embryos developing to live fetuses after transfer to recipients.
- 日本哺乳動物卵子学会の論文
日本哺乳動物卵子学会 | 論文
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