Most early developmental data are lost in bovine embryo culture systems. We developed and validated a method for culture of bovine embryos in groups that allow individual assessment. An autoclavable low-cost multiembryo chamber (MEC) was prepared using a polyester mesh fixed to a glass coverslip. Embryonic development was not affected by MEC. Compared to conventional bovine culture system (oil-covered drops, control), cleavage (C, 71.2 ± 7.8%; MEC, 74.3 ± 6.0%), blastocyst rate (C, 29.9 ± 4.4%; MEC, 28.3 ± 5.0%) and blastocyst cell number (C, 94.1 ± 9.7; MEC, 92.9 ± 5.3) were similar. Caspase 3 positive cell index in blastocysts was increased in MEC group, but apoptosis rate was below 5% (C, 2.9 ± 0.5; MEC, 4.6 ± 0.6). Using MEC, we performed a retrospective analysis for ‘failure’ and ‘success’ embryos, based on their ability to reach the blastocyst stage. We detected the majority of ‘success’ embryos displayed 8 cells at 48 h post-insemination (hpi) (48.7%), but blastocysts derived from this pattern presented lower cell numbers (91.3 ± 4.2 vs. 107.9 ± 4.9) and higher apoptosis index (6.2 ± 0.6 vs. 4.4 ± 0.5) than blastocysts from 4-cell embryos at 48 hpi. Most (72.0%) embryos that were at morula stage 120 hpi reached blastocyst stage at 168 hpi. Those blastocysts presented more number of cells than blastocysts derived from embryos exhibiting 16 cells at 120 hpi (108.6 ± 4.1 vs. 83.9 ± 4.8). Combination of embryo kinetics data at 48 and 120 hpi revealed high chances of blastocyst formation for patterns: 8 cells/morula, 4 cells/morula, 8 cells/16 cells and 4 cells/16 cells. Blastocysts formed from 4-cell/morula and 8-cell/morula patterns represented 69% of all 168 hpi blastocysts. Blastocysts derived from 4 cells/16 cells displayed decreased apoptosis (3.1 ± 0.6). Our results suggest that MEC can be used for bovine embryo culture without detrimental effects on development and can help to predict blastocyst formation and quality of in vitro fertilization (IVF) embryos.

Abbreviations: BSA: bovine serum albumine; COC: cumulus–oocyte complex; FERT-TALP: Tyrode’s albumin lactate pyruvate fertilization; FBS: fetal bovine serum; IVF: in vitro fertilization; MEC: multiembryo chamber; PBS: phosphate buffered saline; SOF-AA: synthetic oviductal fluid with amino acids medium; TCM: Tissue Culture Medium

大多数早期发育数据在牛胚胎培养系统中丢失。我们开发并验证了一种可进行个体评估的成群培养牛胚胎的方法。使用固定在玻璃盖玻片上的聚酯网，准备了可高压灭菌的低成本多胚室（MEC）。胚胎发育不受MEC影响。与传统的牛培养系统（油浸滴，对照）相比，卵裂（C，71.2±7.8％; MEC，74.3±6.0％），胚泡率（C，29.9±4.4％; MEC，28.3±5.0％）和胚泡细胞数（C，94.1±9.7； MEC，92.9±5.3）相似。MEC组胚泡中胱天蛋白酶3阳性细胞指数升高，但凋亡率低于5％（C，2.9±0.5； MEC，4.6±0.6）。使用MEC，我们对“失败”和“成功”的胚胎进行了回顾性分析，根据他们达到胚泡阶段的能力而定。我们检测到大多数“成功”的胚胎在授精后48 h（hpi）展示了8个细胞（48.7％），但是从这种模式衍生的胚泡细胞数量较少（91.3±4.2 vs. 107.9±4.9）和较高的细胞凋亡指数（6.2±0.6 vs. 4.4±0.5）比48 hpi时来自4细胞胚胎的胚泡高。处于桑ula期120 hpi的大多数（72.0％）胚胎在168 hpi达到胚泡期。这些胚泡比在120 hpi时具有16个细胞的胚胎胚泡呈现更多的细胞数量（108.6±4.1对83.9±4.8）。胚胎动力学数据在48和120 hpi的组合显示胚泡形成的几率很高：8个细胞/桑ula，4个细胞/桑,、 8个细胞/ 16个细胞和4个细胞/ 16个细胞。由4细胞/桑ula和8细胞/桑ula形成的囊胚占所有168 hpi囊胚的69％。来自4个细胞/ 16个细胞的胚泡显示出降低的凋亡（3.1±0.6）。我们的结果表明，MEC可用于牛胚胎培养，而对发育无不利影响，并可帮助预测胚泡的形成和质量。体外受精（IVF）胚胎。

缩写： BSA：牛血清白蛋白；COC：卵丘-卵母细胞复合体；FERT-TALP：蒂罗德的白蛋白乳酸丙酮酸受精；FBS：胎牛血清；试管婴儿：体外受精；MEC：多胚室；PBS：磷酸盐缓冲盐水；SOF-AA：含氨基酸的合成输卵管液；中医：组织培养基