School of Veterinary Medicine, Department of Animal Health, UNESP, Rua Clóvis Pestana, 793, 16050-680, Araçatuba, São Paulo, Brazil.
In Vitro Cell Dev Biol Anim. 2011 Jun;47(5-6):361-7. doi: 10.1007/s11626-011-9400-0. Epub 2011 Mar 18.
Aiming to standardize in vitro production of bovine embryos and to obtain supplements to replace serum in culture media, this study evaluated the nuclear maturation kinetics and embryonic development in bovine after in vitro maturation (IVM) and culture (IVC) with several macromolecules (animal origin: bovine serum albumin (BSA), fetal calf serum (FCS); synthetic: polyvinyl alcohol (PVA), polyvinyl pyrrolidone (PVP), Ficoll, and Knockout) at two oxygen tensions (20% and 5% O(2)). Regarding nuclear kinetics, neither the presence of the expected stage (metaphase I, transition anaphase to telophase, and metaphase II) at each evaluation moment (6, 18, and 24 h after IVM, respectively) nor the accelerated polar body emission (at 18 h after IVM) related developmental competence to blastocyst stage when different supplements were compared. Independently of supplement, cleavage rates at 20% O(2) (61.6-79.2%) were higher than at 5% O(2) (38.9-58.7%). At 20% O(2), higher blastocyst and hatching rates, respectively, were obtained in treatments BSA, FCS, Knockout, and control group (IVM with FCS and IVC with BSA + FCS, 14.0-23.5% and 6.8-15.4%) in comparison to PVA, PVP, and Ficoll (0%). The same was observed at 5% O(2) for blastocyst rates with BSA, FCS, Knockout, and control (5.4-16.8%) and for hatching rates with BSA, FCS, and control (2.0-11.1%). We can conclude that producing bovine embryos at 20% O(2) during the entire IVP process resulted in higher developmental rates than at 5% O(2). In addition, while defined macromolecules PVA, PVP, and Ficoll were not suitable for embryonic development, the synthetic serum Knockout was able to replace serum and albumin for IVP in bovine at 20% O(2).
本研究旨在规范牛胚胎的体外生产,并获得替代培养基中血清的补充物,因此评估了在两种氧张力(20%和 5%O 2 )下,几种大分子(动物来源:牛血清白蛋白(BSA),胎牛血清(FCS);合成:聚乙烯醇(PVA),聚乙烯吡咯烷酮(PVP),Ficoll 和 Knockout)对牛体外成熟(IVM)和体外培养(IVC)后的核成熟动力学和胚胎发育的影响。关于核成熟动力学,在每个评估时刻(分别为 IVM 后 6、18 和 24 小时),均未观察到预期阶段(中期 I,从后期向末期的过渡和中期 II)的存在,也未观察到极体加速排放(在 IVM 后 18 小时)与不同补充物相关的囊胚阶段的发育能力。无论补充物如何,在 20%O 2 时的卵裂率(61.6-79.2%)均高于在 5%O 2 时的卵裂率(38.9-58.7%)。在 20%O 2 下,与 PVA、PVP 和 Ficoll(0%)相比,BSA、FCS、Knockout 和对照组(用 FCS 进行 IVM 并用 BSA+FCS 进行 IVC)分别获得更高的囊胚和孵化率,分别为 14.0-23.5%和 6.8-15.4%。在 5%O 2 下,BSA、FCS、Knockout 和对照组的囊胚率和 BSA、FCS 和对照组的孵化率也观察到相同的结果(分别为 5.4-16.8%和 2.0-11.1%)。我们可以得出结论,在整个 IVP 过程中,在 20%O 2 下生产牛胚胎比在 5%O 2 下产生更高的发育率。此外,虽然定义明确的大分子 PVA、PVP 和 Ficoll 不适合胚胎发育,但合成血清 Knockout 能够在 20%O 2 下替代牛的 IVP 中的血清和白蛋白。