Generation of porcine diploid blastocysts after injection of spermatozoa grown in nude mice
Generation of porcine diploid blastocysts after injection of spermatozoa grown in nude mice
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DOI：
10.1016/j.theriogenology.2008.10.020
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发表时间：
2009-07-01
期刊：
影响因子：
2.8
通讯作者：
Kikuchi, K.
中科院分区：
文献类型：
--
作者：
Nakai, M.;Kaneko, H.;Kikuchi, K.
预计通过睾丸异种移植利用精原细胞将为雄性配子的保护开辟新的途径。为了建立这一新的猪遗传保存技术，我们将其与卵胞浆内单精子注射（ICSI）相结合。将来自新生仔猪的睾丸组织移植到去势裸鼠的背部皮肤下，所述睾丸组织含有仅由生殖母细胞/精原细胞组成的生精索。异种移植后125和192天之间，从65只宿主小鼠中的41只（63.1%）回收精子（形态学上类似于附睾精子）。成年公猪的睾丸精子用作阳性对照。将单个精子注射到体外成熟的猪卵母细胞中，并对卵母细胞进行电刺激和培养（分别为移植-ICSI和睾丸-ICSI）。两个ICSI组的囊胚率（分别为24.9%和37.4%）高于（P < 0.05）未注射程序（孤雌生殖; 12.7%）和注射少量注射缓冲液后（假手术; 13.0%）。移植物-ICSI组和睾丸-ICSI组的二倍体囊胚率（48.9%和60.6%）均高于孤雌激活组和假手术组（13.5%和28.0%）（P < 0.05）。因此，我们证明了注射异种精子的猪卵母细胞具有体外发育到囊胚阶段的能力。(C)2009爱思唯尔公司All rights reserved.
It is anticipated that the utilization of spermatogonia through testicular xenografting will open new avenues for the conservation of male gametes. With the aim of establishing this new technique for genetic preservation of pigs, we used it in combination with intracytoplasmic sperm injection (ICSI). Testicular tissues derived from neonatal piglets, which contained seminiferous cords consisting of only gonocytes/spermatogonia, were transplanted under the back skin of castrated nude mice. Between 125 and 192 d after xenografting, sperm (morphologically similar to epididymal sperm) were recovered from 41 of the 65 host mice (63.1%). Testicular spermatozoa from adult boars were used as a positive control. A single spermatozoon was injected into an in vitro matured porcine oocyte, and the oocytes were electro-stimulated and cultured (graft-ICSI and testis-ICSI, respectively). Blastocyst rates in both ICSI groups (24.9% and 37.4%, respectively) were higher (P < 0.05) than those without the injection procedure (parthenogenetic; 12.7%) and after injection of a small amount of injection buffer (sham; 13.0%). Rates of diploid blastocysts in both graft-ICSI and testis-ICSI groups (48.9% and 60.6%) were higher (P < 0.05) than those in the parthenogenetic and sham groups (13.5% and 28.0%). Therefore, we demonstrated that porcine oocytes injected with xenogeneic sperm have in vitro developmental ability to the blastocyst stage. (C) 2009 Elsevier Inc. All rights reserved.