Cattle production by intracytoplasmic sperm injection into oocytes vitrified after ovum pick-up
Cattle production by intracytoplasmic sperm injection into oocytes vitrified after ovum pick-up
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DOI：
10.1016/j.theriogenology.2022.03.022
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发表时间：
2022-04-06
期刊：
影响因子：
2.8
通讯作者：
Kawahara, Manabu
中科院分区：
文献类型：
--
作者：
Kagawa, Shinjiro;Hiraizumi, Shingo;Kawahara, Manabu
胞浆内单精子注射 (ICSI)、取卵后卵母细胞玻璃化冷冻 (OPU) 和体外成熟等生殖技术在高效养牛方面具有令人难以置信的潜力。然而，使用玻璃化 OPU 卵母细胞通过 ICSI 产生的胚胎的发育能力仍然未知。在这里，我们的目的是评估这些胚胎从早期胚胎期到足月的发育能力。使用玻璃化 OPU 卵母细胞的 ICSI 胚胎在体外培养过程中的卵裂率显着低于使用新鲜 OPU 卵母细胞的对照体外受精 (IVF) 胚胎 (30.9 +/- 4.5% vs. 65.9 +/- 7.0%) (P < 0.05)，但囊胚与卵裂胚胎的比例显着高于使用玻璃化卵母细胞的 IVF 胚胎。 OPU 卵母细胞（55.9 +/- 10.8% 对比 23.2 +/- 9.3%）（P < 0.05）。为了进一步研究使用玻璃化 OPU 卵母细胞的 ICSI 胚胎中细胞分化相关基因的转录水平，通过定量逆转录 PCR 分析了 mRNA（OCT4、NANOG、SOX2、CDX2、GATA3 和 IFNT）的相对丰度。使用玻璃化 OPU 卵母细胞的 ICSI 胚胎和对照 IVF 胚胎之间的表达水平没有显着差异。最后，通过胚胎移植检查使用玻璃化 OPU 卵母细胞的 ICSI 胚胎的足月发育能力，并诞生了两只健康的小牛。这些发现证实，ICSI 和玻璃化冷冻会降低体外发育速率，但这两种程序都可以导致牛胚胎的足月发育。这些结果表明，使用玻璃化 OPU 卵母细胞的 ICSI 胚胎对于牛生产是可行的。(c) 2022 Elsevier Inc. 保留所有权利。
Intracytoplasmic sperm injection (ICSI), oocyte vitrification after ovum pick-up (OPU), and in vitro maturation are reproductive technologies with incredible potential for efficient cattle production. However, the developmental competence of embryos produced by ICSI using vitrified OPU oocytes remains unknown. Here, we aimed to evaluate the developmental competence of these embryos from the early embryo period to full term. The cleavage rate in the ICSI embryos using vitrified OPU oocytes during in vitro culture was significantly lower than those in control in vitro fertilized (IVF) embryos using fresh OPU oocytes (30.9 +/- 4.5% v.s. 65.9 +/- 7.0%) (P < 0.05), but the proportion of blastocysts to cleaved embryos was significantly higher than those of IVF embryos using vitrified OPU oocytes (55.9 +/- 10.8% v.s. 23.2 +/- 9.3%) (P < 0.05). To further investigate the transcription levels of genes related to cell differentiation in ICSI embryos using vitrified OPU oocytes, the relative abundance of mRNAs (OCT4, NANOG, SOX2, CDX2, GATA3, and IFNT) was analyzed by quantitative reverse-transcription PCR. There were no significant differences in the expression levels between ICSI embryos using vitrified OPU oocytes and control IVF embryos. Finally, developmental competence to term in ICSI embryos using vitrified OPU oocytes was examined by embryo transfer, and two healthy calves were born. These findings confirmed that ICSI and vitrification decrease developmental rates in vitro, but both procedures can lead to full-term development of bovine embryos. These results demonstrate that ICSI embryos using vitrification OPU oocytes are viable for cattle production.(c) 2022 Elsevier Inc. All rights reserved.