Beneficial effects of culturing rabbit zygotes to blastocysts in 5% oxygen and 10% carbon dioxide.
Beneficial effects of culturing rabbit zygotes to blastocysts in 5% oxygen and 10% carbon dioxide.
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DOI：
10.1530/jrf.0.1030127
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
P. Farrell;R. Foote
中科院分区：
文献类型：
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作者：
P. Farrell;R. Foote
在注射LH和授精后19 h，从超数排卵的荷兰带兔中收集受精卵。丢弃似乎未受精的卵母细胞。在所有培养处理中，受精卵在每只供体雌性中均匀分布。培养皿含有500微升无大分子的RD培养基，其由等份的RPMI 1640和低葡萄糖Dulbecco改良的Eagle培养基组成。胚胎在39 ℃下在N2加以下气体的几种组合中培养：（1）1%O2：10%CO2;（2）5%O2：10%CO2;和（3）20%O2：10%CO2。对照（4）为95%空气：5% CO2。用来自11个供体的胚胎重复实验，提供295个可用的受精卵。培养84 h后，处理1 ~ 4的囊胚形成率分别为13、86、82和59（P < 0.01）。相应的平均细胞数，包括所有培养的卵裂胚（不包括退化胚），分别为55、183、118和68（P < 0.01）。这些结果表明，10%CO2与5%O2的组合是在合成培养基中培养兔受精卵的更有效的气相，而不是通常使用的5%CO2，并且5%O2上级1%或20%O2。
Zygotes were collected from superovulated Dutch-belted rabbits 19 h after injection of LH and insemination. Oocytes that appeared to be unfertilized were discarded. The zygotes were distributed equally within each donor female across all culture treatments. Culture dishes contained 500 microliters of macromolecule-free RD medium consisting of equal parts of RPMI 1640 and low glucose Dulbecco's modified Eagle's medium. Embryos were cultured at 39 degrees C in several gas combinations of N2 plus the following: (1) 1% O2:10% CO2; (2) 5% O2:10% CO2; and (3) 20% O2:10% CO2. The control (4) was 95% air:5% CO2. The experiment was replicated with embryos from 11 donors providing 295 usable zygotes. After 84 h of culture, the percentages of blastocysts formed in treatments 1 to 4, respectively, were 13, 86, 82 and 59 (P < 0.01). The corresponding mean cell counts, including all cleaved embryos cultured (but not degenerate ones), were 55, 183, 118 and 68 (P < 0.01). These results indicate that 10% CO2 combined with 5% O2 is a more effective gas phase for culturing rabbit zygotes in a synthetic medium than is the commonly used 5% CO2, and that 5% O2 is superior to either 1% or 20% O2.