Embryo production by intracytoplasmic injection of sperm retrieved from Meishan neonatal testicular tissue cryopreserved and grafted into nude mice
Embryo production by intracytoplasmic injection of sperm retrieved from Meishan neonatal testicular tissue cryopreserved and grafted into nude mice
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DOI：
10.1111/asj.13138
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发表时间：
2019-02-01
影响因子：
2
通讯作者：
Noguchi, Junko
中科院分区：
文献类型：
--
作者：
Kaneko, Hiroyuki;Kikuchi, Kazuhiro;Noguchi, Junko
睾丸异种移植与冷冻保存相结合，可以从新生猪的睾丸中回收生殖细胞，从而帮助保存地方猪的遗传多样性。以眉山公猪为例，观察睾丸组织在冷冻保存后移植到裸鼠体内后是否具有生精能力(MS组)。为了进行比较，使用了新生西方杂交雄性仔猪的睾丸组织(WC组)。异种移植后60天(第0天=移植)，MS移植物已经发育出含有精子的生精小管，而WC移植物在第120天首次出现精子。90~120日龄时，MS组小管内含精子细胞和精子的比例高于WC组。此外，在体外成熟的猪卵母细胞中，注射从180d的MS组获得的单个精子发育到囊胚期。注射异种精子后的囊胚形成率为14.6%，而不注射异种精子的囊胚形成率(可归因于孤雌生殖)为6.7%。因此，冷冻保存的眉山睾丸组织在宿主小鼠中获得生精活性的时间比西方杂交小鼠早60天。这种异种精子很可能能够在体外产生胚泡。
Testicular xenografting, combined with cryopreservation can assist conservation of the genetic diversity of indigenous pigs by salvaging germ cells from their neonatal testes. Using Meishan male piglets as an example, we examined whether testicular tissue would acquire the ability to produce sperm after cryopreservation and grafting into nude mice (MS group). For comparison, testicular tissue from neonatal Western crossbreed male piglets was used (WC group). Sixty days after xenografting (day 0 = grafting), MS grafts had already developed seminiferous tubules containing sperm, whereas in the WC grafts, sperm first appeared on day 120. The proportion of tubules containing spermatids and sperm was higher in the MS group than in the WC group between days 90 and 120. Moreover, in vitro-matured porcine oocytes injected with a single sperm obtained from the MS group on day 180 developed to the blastocyst stage. The blastocyst formation rate after injection of the xenogeneic sperm was 14.6%, whereas the ratio in the absence of such injection (attributable to parthenogenesis) was 6.7%. Thus, cryopreserved Meishan testicular tissue acquired spermatogenic activity in host mice 60 days earlier than Western crossbreed tissue. Such xenogeneic sperm are likely capable of generating blastocysts in vitro.