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In vitro canine oocyte nuclear maturation in homologous oviductal cell co-culture with hormone-supplemented media

机译:荷尔蒙补充培养基在同种输卵管细胞中体外培养犬卵母细胞核成熟

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The aim of the present research was to verify the influence of oviductal cell co-culture previously supplemented with steroids (estrogen, progesterone, or both) on IVM rates for oocytes from anestrous bitches that were cultured in vitro for 48, 72 and 96h. Oocytes harvested from anestrous bitches were selected and allocated into four groups: Group 1 (co-culture in oviductal epithelial cells without hormonal supplementation-control); Group 2 (estrogen supplementation); Group 3 (progesterone supplementation); Group 4 (estrogen+progesterone supplementation). The oviductal epithelial cell culture was established 72h prior to oocyte co-culture. After periods of 48, 72 and 96h, the degree of oocyte nuclear maturation was assessed. Co-culture in oviductal epithelial cells with estrogen was not as beneficial for canine IVM as supplementation with progesterone and estrogen, or progesterone supplementation alone. Therefore, it was feasible to use co-culture with oviductal epithelial cells obtained from anestrous bitches for IVM (monolayer culture with oviduct cells previously supplemented with progesterone). Final stages of oocyte maturation were achieved at 72 and 96h of culture; therefore, the duration of maturation for oocytes obtained from bitches in different stages of the estrous cycle should be taken into account.
机译:本研究的目的是验证先前补充了类固醇(雌激素,孕激素或两者)的输卵管细胞共培养对离体母犬的卵母细胞IVM速率的影响,该母体在体外培养48、72和96h。选择从动情雌性处收集的卵母细胞,并分为四组:第一组(在没有荷尔蒙补充控制的输卵管上皮细胞中共培养);第2组(补充雌激素);第三组(补充孕激素);第4组(补充雌激素+孕激素)。在卵母细胞共培养之前72小时建立输卵管上皮细胞培养。在48、72和96小时后,评估卵母细胞核成熟度。与雌激素一起在输卵管上皮细胞中共培养对犬IVM的好处不如补充黄体酮和雌激素或单独补充黄体酮。因此,与从动情母犬获得的输卵管上皮细胞共培养用于IVM是可行的(与先前补充黄体酮的输卵管细胞进行单层培养)。卵母细胞成熟的最后阶段是在培养72和96h时完成的。因此,应该考虑从动情周期不同阶段的母犬获得的卵母细胞的成熟持续时间。

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