Akagi Satoshi, Hosoe Misa, Matsukawa Kazutsugu, Ichikawa Akihiko, Tanikawa Tamio, Takahashi Seiya
Reproductive Biology and Technology Research Team, National Institute of Livestock and Grassland Science, Ibaraki, Japan.
J Reprod Dev. 2010 Aug;56(4):475-9. doi: 10.1262/jrd.09-213h. Epub 2010 May 19.
We fabricated a polydimethylsiloxane (PDMS)-based microwell plate (PDMS-MP) containing 100 microwells with a rounded bottom and examined whether it can be used for culture of individual in vitro fertilized (IVF) embryos or parthenogenetically activated zona-free embryos in cattle. In Experiment 1, we examined the in vitro developmental ability of IVF embryos cultured individually on PDMS-MP. After IVF, 20 embryos were transferred into 100 microl drops on PDMS-MP and cultured individually in each well of PDMS-MP (PDMS group). After 7 days of culture, the embryos in the PDMS group developed to the blastocyst stage at the same rate of those in the control group cultured in a group of 20 embryos without PDMS-MP. There were no differences in total number of cells and the ratio of inner cell mass to total cells between the PDMS and control groups. In Experiment 2, we examined the in vitro developmental ability of parthenogenetically activated zona-free bovine embryos cultured individually on PDMS-MP. The zona-free embryos were cultured individually in each well of a PDMS-MP or in each well produced by pressing a darning needle onto the bottom of a culture dish (WOW group). After 7 days of culture, the blastocyst formation rate and cell number of blastocysts in the PDMS group did not differ from those of the zona-intact embryos in the control group. Also, there were no differences in the blastocyst formation rate and cell number of blastocysts between the WOW and PDMS groups. These results suggest that the culture system using PDMS-MP is useful for individual embryos or zona-free embryos in cattle.
我们制作了一种基于聚二甲基硅氧烷（PDMS）的微孔板（PDMS-MP），其包含100个底部为圆形的微孔，并研究了它是否可用于牛体外受精（IVF）胚胎或孤雌生殖激活的无透明带胚胎的个体培养。在实验1中，我们检测了在PDMS-MP上单独培养的IVF胚胎的体外发育能力。体外受精后，将20个胚胎转移到PDMS-MP上的100微升液滴中，并在PDMS-MP的每个孔中单独培养（PDMS组）。培养7天后，PDMS组的胚胎发育到囊胚阶段的速率与在无PDMS-MP的情况下以20个胚胎一组培养的对照组相同。PDMS组和对照组之间的细胞总数以及内细胞团与总细胞的比例没有差异。在实验2中，我们检测了在PDMS-MP上单独培养的孤雌生殖激活的无透明带牛胚胎的体外发育能力。无透明带胚胎在PDMS-MP的每个孔中或通过将缝衣针压在培养皿底部产生的每个孔中单独培养（WOW组）。培养7天后，PDMS组的囊胚形成率和囊胚细胞数与对照组中有透明带的胚胎没有差异。此外，WOW组和PDMS组之间的囊胚形成率和囊胚细胞数也没有差异。这些结果表明，使用PDMS-MP的培养系统对牛的个体胚胎或无透明带胚胎是有用的。
