Normal reproductive development of pigs produced using sperm retrieved from immature testicular tissue cryopreserved and grafted into nude mice
Normal reproductive development of pigs produced using sperm retrieved from immature testicular tissue cryopreserved and grafted into nude mice
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DOI：
10.1016/j.theriogenology.2014.04.012
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发表时间：
2014-07-15
期刊：
影响因子：
2.8
通讯作者：
Kashiwazaki, Naomi
中科院分区：
文献类型：
--
作者：
Kaneko, Hiroyuki;Kikuchi, Kazuhiro;Kashiwazaki, Naomi
未成熟睾丸组织异种移植结合冷冻保存可以保存和利用青春期前动物的遗传信息。为了建立这种新方法，有必要澄清在携带冷冻异种移植物的宿主小鼠中生长的精子所产生的后代是否表现出正常的生殖发育。本研究检查了猪性成熟过程中性成熟期间性腺激素的血清谱，这些猪是通过使用冷冻异种移植物（CryoXeno 猪；三只雄性和三只雌性）衍生的精子进行胞浆内单精子注射而产生的。我们还通过将雄性 CryoXeno 猪与传统生产的猪交配，并检查其精子的体外受精能力，评估了它们的繁殖能力。对于雌性 CryoXeno 猪，通过使用传统公猪的精液进行人工授精来评估繁殖能力。在雄性 CryoXeno 猪的生长过程中，抑制素和睾酮的血清浓度与常规猪 (n = 4) 表现出相似的变化 (P > 0.17)。睾丸的组织学分析显示 CryoXeno 和传统猪之间生精小管的生长和分化没有差异 (P > 0.2)。三头传统母猪与三头 CryoXeno 雄性交配后产下 13.0 +/- 1.0（平均值的平均标准误差）活仔猪。所有 CryoXeno 猪获得的精子均具有穿透卵母细胞的能力，并且这些受精卵母细胞在体外达到囊胚阶段。在雌性 CryoXeno 猪的生长过程中，血清抑制素谱与传统猪 (n = 5) 中观察到的相似 (P > 0.17)。 CryoXeno 猪的血清孕酮浓度首次上升至 2 ng/mL 以上，时间为 32.0 +/- 2.3 周龄，而传统猪则为 32.0 +/- 3.3 周，这表明两只猪在相似的年龄进入青春期。青春期后，雌性 CryoXeno 猪在使用传统公猪的精液进行人工授精后产下 8.3 +/- 1.7（平均值 +/- 平均值标准误差；n = 3）头活仔猪。总之，这些发现表明雄性和雌性 CryoXeno 猪都具有正常的繁殖能力。 (C) 2014 Elsevier Inc. 保留所有权利。
Xenografting of immature testicular tissue combined with cryopreservation can preserve and use genetic information of prepubertal animals. For establishment of this new approach, it is essential to clarify whether offspring derived from sperm grown in host mice harboring cryopreserved xenografts show normal reproductive development. This study examined serum profiles of gonadal hormones during sexual maturation in pigs generated by intracytoplasmic sperm injection using sperm derived from cryopreserved xenografts (CryoXeno pigs; three males and three females). We also assessed the reproductive abilities of the male CryoXeno pigs by mating them with conventionally produced (conventional) pigs, and by examining the in vitro fertilizing ability of their sperm. For female CryoXeno pigs, reproductive ability was evaluated by artificial insemination with semen from a conventional boar. During the growth of male CryoXeno pigs, the serum concentrations of inhibin and testosterone showed similar changes (P > 0.17) to those in conventional pigs (n = 4). Histologic analyses of the testes revealed no differences (P > 0.2) in the growth and differentiation of seminiferous tubules between CryoXeno and conventional pigs. Three conventional sows delivered 13.0 +/- 1.0 (mean standard error of the mean) live piglets after being mated with the three CryoXeno males. Sperm obtained from all CryoXeno pigs had the ability to penetrate oocytes, and these fertilized oocytes reached the blastocyst stage in vitro. During the growth of female CryoXeno pigs, the serum inhibin profile was similar (P > 0.17) to that observed in conventional pigs (n = 5). The first rise in serum progesterone concentration to more than 2 ng/mL was noted at 32.0 +/- 2.3 weeks of age in the CryoXeno pigs and at 32.0 +/- 3.3 weeks in the conventional pigs, suggesting that both pigs reached puberty at a similar age. After puberty, female CryoXeno pigs farrowed 8.3 +/- 1.7 (mean +/- standard error of the mean; n = 3) live piglets after artificial insemination with semen from a conventional boar. In conclusion, these findings demonstrate that both male and female CryoXeno pigs have normal reproductive abilities. (C) 2014 Elsevier Inc. All rights reserved.