Watanabe Hideaki, Matsumoto Takuya, Nishi Moeka, Kusakabe Ken Takeshi, Kuraishi Takeshi, Hattori Shosaku, Matsumoto Hiromichi, Fukui Emiko, Kuwahata Akiko, Ochi Masanori, Kiso Yasuo, Kai Chieko, Yoshizawa Midori
United Graduate School of Agricultural Science Tokyo University of Agriculture and Technology 183-8509 Fuchu-shi Japan.
Graduate School of Agricultural Science Utsunomiya University 321-8505 Tochigi Japan.
Reprod Med Biol. 2015 Dec 16;15(3):183-186. doi: 10.1007/s12522-015-0229-1. eCollection 2016 Jul.
We explored the possibility of employing intracytoplasmic sperm injection (ICSI), involving oocytes and sperm of owl monkeys, to increase the availability of this species for investigations relating to malaria, etc., by increasing the number of animals in our laboratory.
Two owl monkeys (a female and a male), raised at the Amami Laboratory of the University of Tokyo, were used. Follicular oocytes surrounded with cumulus cells were cultured in vitro for approximately 25 h and cumulus cells were removed with 0.1 % hyaluronidase. Because of the poor motility of caudal epididymal sperm, sperm were injected without adding polyvinylpyrrolidone to immobilize them. The ICSI procedure was performed by an individual with considerable experience of human ICSI.
We were able to produce two owl monkey embryos using ICSI of oocytes that matured to MII stage. Both embryos reached the 10-cell stage at 98 h after ICSI and showed signs of compaction, but failed to cleave further.
Although we successfully produced owl monkey embryos after ICSI, the embryos did not develop to the blastocyst stage. Many parameters need to be studied further, including superovulation, selection of culture media, and selection of good quality sperm in order to achieve successful ICSI in the owl monkey.
我们探讨了采用胞浆内单精子注射(ICSI)技术,利用夜猴的卵母细胞和精子,通过增加我们实验室的动物数量,来提高该物种用于疟疾等相关研究的可用性。
使用了两只在东京大学奄美实验室饲养的夜猴(一只雌性和一只雄性)。将带有卵丘细胞的卵泡卵母细胞在体外培养约25小时,并用0.1%的透明质酸酶去除卵丘细胞。由于附睾尾部精子活力较差,在注射精子时未添加聚乙烯吡咯烷酮来使其固定。ICSI操作由一位有丰富人类ICSI经验的人员进行。
我们利用成熟至MII期的卵母细胞进行ICSI成功培育出了两个夜猴胚胎。两个胚胎在ICSI后98小时均发育至10细胞阶段并出现致密化迹象,但未能进一步分裂。
尽管我们在ICSI后成功培育出了夜猴胚胎,但胚胎未发育至囊胚阶段。为了在夜猴中实现成功的ICSI,许多参数需要进一步研究,包括超排卵、培养基的选择以及优质精子的选择。