Carolan C, Lonergan P, Van Langendonckt A, Mermillod P
Station de Physiologie de la Reproduction, INRA, 37380 Nouzilly, France.
Theriogenology. 1995 Apr 15;43(6):1115-28. doi: 10.1016/0093-691x(95)00075-j.
Employing a total of 3465 bovine oocytes this study was aimed at improving the efficiency of bovine embryo production under defined and undefined conditions. Following in vitro maturation (IVM) and in vitro fertilization (IVF), oocytes were allocated to various culture treatments using synthetic oviduct fluid (SOF). In our 3 experiments we showed that: 1) the addition of fetal calf serum (FCS 10% v/v) to SOF droplets after 20 to 24 h significantly improved blastocyst yields on Day 6 (21 vs 12%; P < 0.01), but not at later stages and resulted in significantly higher Day-8 blastocyst cell numbers (148 +/- 61 vs 92 +/- 35; P < 0.05); 2) the removal of bovine serum albumin (BSA) from the standard SOF medium resulted in significantly reduced blastocyst yields on Days 6, 7 and 8, respectively (17 vs 8%; 28 vs 18%; 31 vs 21%; P < 0.05); 3) the presence or absence of cumulus cells surrounding the presumptive zygote in culture in SOF had no effect on cleavage rate, percentage of 5-8 cell embryos or blastocyst yields (Day 6,7 or 8); 4) the culture of presumptive zygotes in SOF in an atmosphere of 5% CO2 in air (20% O2) resulted in significantly reduced development compared with culture in 5% CO2, 5% O2, 90% N2 in terms of blastocyst yield on Days 6, 7 and 8 and on Day 8 hatching rate, respectively (5 vs 22%; 9 vs 33%; 13 vs 48%; 50 vs 8%; P < 0.001) and 5) embryo density (1 embryo per 1 or 3 microl SOF) or replacing the culture medium every 48 h had no effect when SOF was supplemented with serum; however, under serum-free conditions, changing of the media resulted in a slightly improved Day-6 blastocyst yield such that renewal of serum-free medium mimicked the effect of serum addition.
本研究共使用了3465枚牛卵母细胞，旨在提高在限定和非限定条件下牛胚胎生产的效率。经过体外成熟（IVM）和体外受精（IVF）后，使用合成输卵管液（SOF）将卵母细胞分配到不同的培养处理中。在我们的3个实验中，我们发现：1）在20至24小时后向SOF液滴中添加胎牛血清（FCS，10% v/v）可显著提高第6天的囊胚率（21%对12%；P < 0.01），但在后期阶段则不然，且导致第8天的囊胚细胞数量显著增加（148±61对92±35；P < 0.05）；2）从标准SOF培养基中去除牛血清白蛋白（BSA）分别导致第6、7和8天的囊胚率显著降低（17%对8%；28%对18%；31%对21%；P < 0.05）；3）在SOF中培养时，假定受精卵周围有无卵丘细胞对卵裂率、5 - 8细胞胚胎百分比或囊胚率（第6、7或8天）没有影响；4）与在5% CO2、5% O2、90% N2的环境中培养相比，在含5% CO2的空气中（20% O2）培养假定受精卵，在第6、7和8天的囊胚率以及第8天的孵化率方面均导致发育显著降低（5%对22%；9%对33%；13%对48%；50%对8%；P < 0.001）；5）当SOF添加血清时，胚胎密度（每1或3微升SOF含1个胚胎）或每48小时更换培养基没有影响；然而，在无血清条件下，更换培养基导致第6天的囊胚率略有提高，使得无血清培养基的更新模拟了添加血清的效果。
