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In vitro embryo production from cumulus-oocyte complexes type II in Bos indicus cattle

机译:来自印度s牛的II型卵-卵母细胞复合物的体外胚胎生产

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The morphological selection of cumulus-oocyte complexes (COCs) is an important step for in vitro embryo production. It has been suggested that COC showing signs of atresia have the ability to generate embryos. The objective of this work was to evaluate the effect of COC morphology from Bos indicus animals with signs of early atresia versus no signs of atresia on in vitro embryo production. COC were classified in: Type I (TI): homogeneous ooplasm with ≥ 4 layers of compact cumulus cells (CC) and Type II (TII): granular ooplasm and ≥ 4 layers of CC slightly expanded. The COC were matured in vitro for 24 hours in TCM199 medium and subsequently fertilized in vitro for 18 h. The suspected zygotes were cultured in vitro for seven days in modified SOFaa medium. Embryonic quality was determined by blastomeric count following staining with Hoechst 33342. Student test was used to determine statistical differences for cleavage, blastocyst rate and blastomeric counts between types of COC. The cleavage rate for TI (n = 220) and TII (n = 161) was 88 ?± 4% and 89 ?± 8% respectively (p 0.05); embryo development rate was 36 ?± 7% and 33 ?± 8% (p0.05) respectively. The blastomeric count for both groups was 101 and 104 cells for TI and TII respectively (n = 10), (p0.05). These results demonstrate that there is no difference in the quantity and quality of embryos produced in vitro using COC type I or type II, suggesting that both types could be used for bovine in vitro embryo production in Bos indicus cows.
机译:卵-卵母细胞复合物(COC)的形态选择是体外胚胎生产的重要步骤。已经表明,显示闭锁迹象的COC具有产生胚胎的能力。这项工作的目的是评估具有早期闭锁征兆与没有闭锁征兆的Bos indicus动物的COC形态对体外胚胎产生的影响。 COC分为:I型(TI):均质卵泡,具有≥4层的致密卵丘细胞(CC),II型(TII):粒状卵质,≥4层CC,略有膨胀。 COC在TCM199培养基中体外成熟24小时,然后在体外受精18 h。将怀疑的受精卵在改良的SOFaa培养基中体外培养7天。胚胎质量由Hoechst 33342染色后的胚泡计数确定。学生测试用于确定COC类型之间卵裂,胚泡率和胚泡计数的统计学差异。 TI(n = 220)和TII(n = 161)的裂解率分别为88±±4%和89±±8%(p> 0.05)。胚胎发育率分别为36±±7%和33±±8%(p> 0.05)。对于TI和TII,两组的胚球计数分别为101和104个细胞(n = 10),(p> 0.05)。这些结果表明,使用I型或II型COC体外生产的胚胎的数量和质量没有差异,这表明这两种类型均可用于印度dic牛的牛体外胚胎生产。

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