Education Program in Reproduction & Development, Department of Obstetrics & Gynaecology, School of Clinical Sciences, Monash University, Clayton, Victoria, Australia.
The Ritchie Centre, Hudson Institute of Medical Research, Clayton, Victoria, Australia.
PLoS One. 2020 Dec 28;15(12):e0244411. doi: 10.1371/journal.pone.0244411. eCollection 2020.
The Egyptian or Common spiny mouse (A. cahirinus) is the first rodent species to show human-like menstruation and spontaneous decidualisation. We consider from these, and its other, human-like characteristics that this species will be a more useful and appropriate small animal model for human reproductive studies. Based on this, there is a need to develop specific laboratory-based assisted reproduction protocols including superovulation, in-vitro fertilisation, embryo cryopreservation and transfer to expand and make this model more relevant. Because standard rodent superovulation has not been successful in the spiny mouse, we have selected to test a human protocol. Female spiny mice will receive a subcutaneous GnRH agonist implant and be allowed to recover. Menstrual cycle lengths will then be allowed to stabilize prior to ovarian stimulation. After recovery, females will be injected IP once a day for 4 days with a FSH analogue, to induce follicular growth, and on day 5 will be injected IP with a hCG analogue to trigger ovulation. Females will either be culled 36hrs after trigger to collect oocytes or immediately paired with a stud male and two cell embryos collected 48hrs later. Mature oocytes will be inseminated using fresh spiny mouse spermatozoa and all in-vitro grown and in-vivo collected two cell embryos will be cryopreserved using methods developed in a close spiny mouse relative, the Mongolian gerbil. For embryo transfer, vitrified embryos will be rapidly warmed and non-surgically transferred to surrogate mice. Surrogates will be monitored until pregnancy is apparent (roughly 30 days) and then left undisturbed until birth, 38-40 days after transfer. By successfully developing robust assisted reproduction protocols in A. cahirinus we will be able to use this rodent as a more effective model for human reproduction.
埃及或普通刺鼠（A. cahirinus）是第一种表现出类似人类月经和自发性蜕膜化的啮齿动物。鉴于这些以及其他类似人类的特征，我们认为该物种将成为更有用和更适合人类生殖研究的小型动物模型。基于此，需要开发特定的基于实验室的辅助生殖协议，包括超排卵、体外受精、胚胎冷冻保存和移植，以扩大和使该模型更相关。由于标准啮齿动物的超排卵在刺鼠中没有成功，我们选择测试一种人类方案。雌性刺鼠将接受皮下 GnRH 激动剂植入并允许恢复。然后允许月经周期长度稳定后再进行卵巢刺激。恢复后，雌性将每天一次通过 IP 注射 FSH 类似物 4 天，以诱导卵泡生长，并在第 5 天通过 IP 注射 hCG 类似物触发排卵。雌性将在触发后 36 小时内被扑杀以收集卵母细胞，或者立即与雄性刺鼠配对，并在 48 小时后收集两细胞胚胎。成熟的卵母细胞将使用新鲜的刺鼠精子进行授精，所有体外培养和体内收集的两细胞胚胎将使用与蒙古沙鼠密切相关的方法进行冷冻保存。对于胚胎移植，将冷冻的胚胎快速解冻并通过非手术方式移植到代孕小鼠中。代孕鼠将被监测，直到怀孕明显（大约 30 天），然后在转移后 38-40 天左右不受干扰，直到分娩。通过在 A. cahirinus 中成功开发强大的辅助生殖协议，我们将能够使用这种啮齿动物作为更有效的人类生殖模型。
