Research for the establishment of artificial indoor breeding systems for the Japanese monkey using developmental technology

1999-2002年对日本猕猴（Macaca fuscata）胚胎移植（ET）与人工授精、体外受精（IVF）、胞浆内单精子注射（ICSI）、体外成熟（IVM）等生殖工程技术相关的研究结果如下：(1)建立了腹腔镜下从eCG（PMSG）、hMG和FSH刺激的卵巢中提取成熟卵子的方法。(2)建立了腹腔镜下2-4个细胞胚胎移植至输卵管的IVF-ET方法。(3)利用该方法在日本猴体内成功实现世界首例IVF-ET妊娠和活产。(4)建立电刺激或直接从安乐死受试者附睾采集精子的液氮冷冻保存方法。(5)建立冷冻精子ICSI-ET方法。利用该方法成功培育出世界上第一个ICSI-ET食蟹猕猴胎儿。(6)建立了zygo…More IVM方法。利用这种方法在日本猴子和食蟹猕猴身上成功建立了世界上第一个胚胎干细胞（ES细胞）。(7)液氮受精卵低温保存取得突破性进展。到今天为止，有9只人工饲养的猪断奶了。截至2002年3月，已确认28例人工受孕。(9)所有人工养殖个体沙门氏菌、志贺氏菌、结核病、内寄生虫、外寄生虫、b病毒、HSV-1、SIV、SRV、SVV、丝状病毒、STLV、麻疹抗体均阴性。生产特定的无病原体（SPF）个体现在是可能的。（10）成功培育出ISCI-ET型日本猴仍然难以捉摸：必须找出原因。现在在日本猴子身上建立了以下方法：利用腹腔镜从受刺激的卵巢中收集卵子；ICSI;腹腔镜下输卵管ET；精子和受精卵冷冻保存。所有必要的准备工作，计划，室内饲养日本猴使用这些方法相信已经完成。少

Results of four year (1999-2002) investigation of embryo transfer (ET) in the Japanese monkey (Macaca fuscata) in association with reproductive engineering techniques such as artificial insemination, in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI), in vitro maturation (IVM), etc., are as follows :(1) Established method for retrieval of mature ova from eCG(PMSG), hMG and FSH stimulated ovaries using laparoscopy.(2) Established IVF-ET method for transfer of 2-4 cell embryo to the fallopian tube under laparoscopic observation.(3) Successfully produced world's first IVF-ET pregnancy and live birth in the Japanese monkey using this method.(4) Established liquid nitrogen cryopreservation method for sperm collected either by electrostimulation or directly from euthanized subjects' epididymis.(5) Established cryopreserved sperm ICSI-ET method. Successfully produced world's first ICSI-ET crab eating macaque (Macaca fascicularis) fetus using this method.(6) Established zygo … More te IVM method. Successfully established world's first embryonic stem cell (ES cell) in the Japanese monkey and the crab eating macaque using this method.(7) Breakthrough in liquid nitrogen zygote cryopreservation close.(8) As of today nine artificially produced individuals weaned. As of March, 2002, 28 artificial impregnations confirmed.(9) All artificially produced individuals confirmed negative for salmonella, shigella, tuberculosis, internal parasites, external parasites, B-virus, HSV-1, SIV, SRV, SVV, filovirus, STLV, and measles antibodies. Production of specific pathogen free (SPF) individual now possible.(10) Success producing a ISCI-ET Japanese monkey remains elusive : must identify reason why.Following methods now established in the Japanese monkey : ova collection from stimulated ovary using laparoscopy ; ICSI ; fallopian tube ET using laparoscopy ; sperm and fertilized zygote cryopreservation. All preparations necessary for planned, indoor breeding of the Japanese monkey using these methods believed complete. Less