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半胱胺、白血病抑制因子和Y27632对山羊卵母细胞体外成熟和胚胎发育的协同作用。

Synergistic effect of cysteamine, leukemia inhibitory factor, and Y27632 on goat oocyte maturation and embryo development in vitro.

作者信息

An Liyou, Liu Jiao, Du Yinyan, Liu Zhihui, Zhang Fenli, Liu Yanhong, Zhu Xiumei, Ling Pingping, Chang Shiwei, Hu Yeshu, Li Yi, Xu Bowen, Yang Lan, Xue Fei, Presicce Giorgio Antonio, Du Fuliang

机构信息

Jiangsu Key Laboratory for Molecular and Medical Biotechnology, College of Life Sciences, Nanjing Normal University, Nanjing 210046, PR China.

Lannuo Biotechnologies Wuxi Inc., Wuxi 214000, PR China.

出版信息

Theriogenology. 2018 Mar 1;108:56-62. doi: 10.1016/j.theriogenology.2017.11.028. Epub 2017 Nov 26.

Abstract

Goat oocyte in vitro maturation is associated with a variable efficiency of embryo development after in vitro fertilization (IVF). Here, we developed a novel maturation procedure to evaluate the cellular effect of cysteamine (Cys), leukemia inhibitory factor (LIF) and Y27632 on oocyte in vitro maturation in native Chinese Yangtze river white goats. Oocytes were collected by slicing ovary tissues and matured for 24 h in vitro prior to IVF. Presumptive fertilized oocytes were cultured in embryo media for 8 days. Maturation rates were similar in gonadotropin basal maturation medium and the same medium supplemented with Cys, LIF, or Y27362 (41.0-48.0%; P > 0.05). However, when two substances were co-supplemented into the medium, the maturation rate was higher in the Cys+LIF group than in the LIF+Y27362 and Cys+Y27362 groups (60.0% vs. 43.1% and 25.8%, respectively; P < 0.05). Co-supplementation of all three substances into the medium achieved the highest maturation rate (67.5%; P < 0.05). Compared with oocytes in gonadotropin basal maturation medium, those in medium supplemented with Cys showed increased fertilization (56.1% vs. 72.1%), cleavage (36.7% vs. 44.8%), and blastocyst development (1.7% vs. 4.2%), respectively (P < 0.05). Cys+LIF supplementation further improved fertilization (81.6%), cleavage (54.9%), and blastocyst development (6%; P < 0.05). Furthermore, combined supplementation of all three substances resulted in the best fertilization (84.9%), cleavage (70.7%), and blastocyst development (10.3%; P < 0.05). Resultant IVF blastocysts possessed an average cell number as high as 276 ± 45 per embryo. This is the first study to report increased efficiency of caprine oocyte maturation by combined Cys, LIF, and Y27632 supplementation into basal maturation medium, leading to improved fertilization and embryo development in vitro post-IVF.

摘要

山羊卵母细胞体外成熟与体外受精(IVF)后胚胎发育的效率变化相关。在此,我们开发了一种新的成熟程序,以评估半胱胺(Cys)、白血病抑制因子(LIF)和Y27632对中国本土长江白山羊卵母细胞体外成熟的细胞效应。通过切割卵巢组织收集卵母细胞,并在IVF前体外成熟24小时。将推定受精的卵母细胞在胚胎培养基中培养8天。在促性腺激素基础成熟培养基以及添加了Cys、LIF或Y27362的相同培养基中,成熟率相似(41.0 - 48.0%;P > 0.05)。然而,当两种物质共同添加到培养基中时,Cys + LIF组的成熟率高于LIF + Y27362组和Cys + Y27362组(分别为60.0%对43.1%和25.8%;P < 0.05)。将所有三种物质共同添加到培养基中成熟率最高(67.5%;P < 0.05)。与促性腺激素基础成熟培养基中的卵母细胞相比,添加Cys的培养基中的卵母细胞受精率(56.1%对72.1%)、卵裂率(36.7%对44.8%)和囊胚发育率(1.7%对4.2%)分别增加(P < 0.05)。添加Cys + LIF进一步提高了受精率(81.6%)、卵裂率(54.9%)和囊胚发育率(6%;P < 0.05)。此外,三种物质联合添加导致最佳的受精率(84.9%)、卵裂率(70.7%)和囊胚发育率(10.3%;P < 0.05)。所得IVF囊胚每个胚胎平均细胞数高达276 ± 45个。这是第一项报道通过在基础成熟培养基中联合添加Cys、LIF和Y27632提高山羊卵母细胞成熟效率,从而导致IVF后体外受精和胚胎发育改善的研究。

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