Lipid accumulation occurs in cultured embryos and is associated with reduced cryotolerance. Here we report the use of a multiple pathway lipid modulator cocktail (l-carnitine, linoleic acid and forskolin) to improve cryosurvival. First, we stained oocytes and embryos with Oil Red to examine the time course of lipid accumulation during in vitro fertilization (IVF) and embryo culture. Then we evaluated the effects of the lipid modulators cocktail on lipid content, developmental rates and survival after vitrification. In our conditions, lipid accumulation was detected (P < 0.05) at the end of in vitro maturation (IVM) and after 4 days of embryo culture (D4-D5). In experiment 1, we used lipid modulator cocktail during IVM. Reduced (P < 0.05) lipid accumulation was detected in oocytes (Control: 49.9 ± 1.6, Lip. Mod. IVM: 45.0 ± 1.8) but no changes were present at blastocyst stage (Control: 62.4 ± 2.6, Lip. Mod. IVM: 66.8 ± 2.7). Treated oocytes presented decreased (P < 0.05) blastocyst rates and lower (P < 0.05) re-expansion after vitrification. In experiment 2, lipid modulators cocktail was used during embryo culture (from D4–D7 or D6–D7). Treatment had an effect on lipid metabolism, as lipid content was increased (P < 0.05) in D7 blastocysts in treated groups (Control: 52.7 ± 3.1a, D4: 65.9 ± 2.6b, D6: 78.1 ± 2.7b). However, no effect was present for cleavage, blastocyst and cryosurvival rates. No difference was detected in mean cell number comparing the three groups (Control: 78.9 ± 9.6, D4: 82.6 ± 16.5, D6: 68.3 ± 7.8), but apoptosis rate was increased (P < 0.05) in vitrified-warmed blastocysts from treated groups (Control: 14.77*, D4: 22.28, D6: 22.22). We concluded that the combined use of lipid modulators was efficient to promote changes in lipid content of oocytes and embryos in bovine, but those changes did not reflect positively on embryo development or cryosurvival.

脂质积累发生在培养的胚胎中，并与低温耐受性降低有关。在这里，我们报告了使用多途径脂质调节剂混合物（左旋肉碱、亚油酸和毛喉素）来提高冷冻存活率。首先，我们用油红对卵母细胞和胚胎进行染色，以检查体外受精(IVF) 和胚胎培养过程中脂质积累的时间过程。然后我们评估了脂质调节剂混合物对脂质含量、发育率和玻璃化后存活率的影响。在我们的条件下，在体外成熟结束 (IVM) 和胚胎培养 4 天后 (D4-D5)检测到脂质积累 ( P < 0.05 )。在实验 1 中，我们在 IVM 期间使用了脂质调节剂鸡尾酒。减少（P < 0.05) 在卵母细胞中检测到脂质积累（对照：49.9 ± 1.6，Lip. Mod. IVM：45.0 ± 1.8），但在囊胚期没有变化（对照：62.4 ± 2.6，Lip. Mod. IVM：66.8 ± 2.7)。经处理的卵母细胞在玻璃化冷冻后囊胚率降低（P < 0.05），再膨胀率降低（P < 0.05）。在实验 2 中，在胚胎培养过程中使用了脂质调节剂混合物（从 D4-D7 或 D6-D7）。治疗对脂质代谢有影响，因为治疗组中 D7 囊胚中的脂质含量增加（P < 0.05）（对照组：52.7 ± 3.1 a，D4：65.9 ± 2.6 b，D6：78.1 ± 2.7 b）。然而，对卵裂、囊胚和冷冻存活率没有影响。与三组相比，平均细胞数未检测到差异（对照组：78.9 ± 9.6，D4：82.6 ± 16.5，D6：68.3 ± 7.8），但处理组的玻璃化加热胚泡细胞凋亡率增加（P < 0.05） （对照：14.77*，D4：22.28，D6：22.22）。我们得出结论，脂质调节剂的联合使用可有效促进牛卵母细胞和胚胎脂质含量的变化，但这些变化并未对胚胎发育或冷冻存活产生积极影响。