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Comparison of the Effects of Oviductal Cell Coculture and Oviductal Cell-conditioned Medium On the Development and Metabolic-activity of Cattle Embryos

机译:输卵管细胞共培养和输卵管条件培养液对牛胚胎发育和代谢活性影响的比较

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摘要

The objective of this study was to compare the development and metabolic activity of cattle embryos co-cultured with bovine oviductal cells or cultured in serum-free medium previously conditioned by bovine oviductal cells. Zygotes were produced by in vitro fertilization of oocytes from bovine ovaries obtained from an abattoir. Development to the four-cell stage occurred by 48 h after fertilization in both culture systems, but co-cultured embryos reached the 16-cell stage by 96 h, whereas those cultured in conditioned medium did not do so until 24 h later. Similarly, the morula and blastocyst stages were reached 24 h earlier in co-culture than in conditioned medium. There were significantly more cells in the blastocysts from co-culture (96.8 +/- 6.1 versus 56.7 +/- 3.3; P less than or equal to 0.0001). The metabolism of glutamine did not differ between embryos cultured in the two systems, but the metabolism of glucose was significantly greater in embryos cultured in conditioned medium. The first significant increase in glucose metabolism occurred between the four-cell and the 16-cell stages in embryos cultured in conditioned medium, but occurred between the 16-cell and morula stages in the co-cultured embryos, such that the glucose metabolism was significantly greater at the 16-cell stage in embryos cultured in conditioned medium compared with co-cultured embryos (6.5 +/- 1.0 versus 2.5 +/- 0.4 pmol per embryo per 3 h, P less than or equal to 0.0001). The concentration of glucose was significantly less, and that of lactate significantly greater, in co-culture medium than in conditioned medium. The results suggest that the activity of enzymes involved in glucose transport or metabolism in the early cattle embryo can be affected by the prior culture conditions, and that a high rate of glucose metabolism may be unfavourable for development.
机译:这项研究的目的是比较与牛输卵管细胞共培养或在事先经牛输卵管细胞调节的无血清培养基中培养的牛胚胎的发育和代谢活性。合子通过体外受精从来自屠宰场的牛卵巢卵母细胞中产生。在两个培养系统中,受精后均在受精后48小时发育到四细胞阶段,但共培养的胚胎在96 h时达到了16细胞阶段,而在条件培养基中培养的胚直到24 h才发育。类似地,共培养比条件培养基早了桑ula和胚泡阶段24小时。共培养的胚泡中有明显更多的细胞(96.8 +/- 6.1对56.7 +/- 3.3; P小于或等于0.0001)。在两种系统中培养的胚之间,谷氨酰胺的代谢没有差异,但是在条件培养基中培养的胚中,葡萄糖的代谢明显更高。葡萄糖代谢的首次显着增加发生在条件培养基中培养的胚胎的四细胞阶段和十六细胞阶段之间,但发生在共培养的胚胎的十六细胞阶段和桑壳阶段之间,因此葡萄糖代谢显着与共培养的胚胎相比,在条件培养基中培养的胚胎在16细胞阶段具有更大的优势(每3h每个胚胎6.5 +/- 1.0对2.5 +/- 0.4 pmol,P小于或等于0.0001)。与条件培养基相比,共培养基中的葡萄糖浓度明显更低,而乳酸的浓度明显更高。结果表明,早期牛胚胎中参与葡萄糖转运或代谢的酶的活性可能受到先前培养条件的影响,并且高葡萄糖代谢率可能不利于发育。

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