Somfai Tamás, Inaba Yasushi, Aikawa Yoshio, Ohtake Masaki, Kobayashi Shuji, Konishi Kazuyuki, Nagai Takashi, Imai Kei
National Livestock Breeding Center, Odakurahara 1, Odakura, Nishigo, Fukushima 961-8511, Japan.
Acta Vet Hung. 2010 Dec;58(4):465-74. doi: 10.1556/AVet.58.2010.4.7.
The aim of the present study was to optimise the culture conditions for the in vitro production of bovine embryos. The development of in vitro fertilised bovine oocytes in CR1aa supplemented with 5% calf serum and IVD101 culture media were compared using traditional microdrops and Well of the Well (WOW) culture systems either under 5% or 20% oxygen tension. After 7 days of culture, a significantly higher blastocyst formation rate was obtained for embryos cultured in CR1aa medium compared to those cultured in IVD101, irrespective of O2 tensions and culture systems. The blastocyst formation in IVD101 was suppressed under 20% O2 compared to 5% O2 . Despite their similar total cell numbers, higher rates of inner cell mass (ICM) cells were observed in blastocysts developed in IVD101 medium than in those developed in CR1aa, irrespective of O2 tensions. There was no significant difference in blastocyst formation, total, ICM and trophectoderm (TE) cell numbers between embryos obtained by microdrop and WOW culture systems irrespective of the culture media and O2 tensions used. In conclusion, CR1aa resulted in higher blastocyst formation rates irrespective of O2 tension, whereas IVD101 supported blastocyst formation only under low O2 levels but enhanced the proliferation of ICM cells.
本研究的目的是优化体外生产牛胚胎的培养条件。使用传统微滴培养系统和“孔中孔”（WOW）培养系统，在5%或20%氧气张力条件下，比较体外受精牛卵母细胞在添加5%小牛血清的CR1aa培养基和IVD101培养基中的发育情况。培养7天后，无论氧气张力和培养系统如何，与在IVD101培养基中培养的胚胎相比，在CR1aa培养基中培养的胚胎获得的囊胚形成率显著更高。与5%氧气张力相比，在20%氧气张力下IVD101中的囊胚形成受到抑制。尽管囊胚的总细胞数相似，但无论氧气张力如何，在IVD101培养基中发育的囊胚中观察到的内细胞团（ICM）细胞比例高于在CR1aa培养基中发育的囊胚。无论使用何种培养基和氧气张力，通过微滴培养系统和WOW培养系统获得的胚胎在囊胚形成、总细胞数、ICM细胞数和滋养外胚层（TE）细胞数方面均无显著差异。总之，无论氧气张力如何，CR1aa都能产生更高的囊胚形成率，而IVD101仅在低氧气水平下支持囊胚形成，但能促进ICM细胞的增殖。
