Medicine and Surgery Department of the Faculty of Veterinary Medicine of Madrid, Universidad Complutense de Madrid, Avd Puerta de Hierro s/n, 28040, Madrid, Spain.
Department of Physiology of the Faculty of Veterinary Medicine, Complutense University o Madrid, Avda. Puerta de Hierro s/n, 28040, Madrid, Spain.
Theriogenology. 2024 Nov;229:47-52. doi: 10.1016/j.theriogenology.2024.08.017. Epub 2024 Aug 13.
Ovum Pick Up (OPU) is a minimally invasive technique widely used in cattle and mares for oocyte retrieval, involving ultrasound-guided puncture of ovarian follicles. It has been demonstrated that this technique is safe for its repeated use in the same female without affecting her reproductive health, allowing for the retrieval of oocytes in individuals regardless of their reproductive status. The oocytes obtained through OPU can subsequently be used for in vitro embryo production (IVP) using assisted reproductive techniques (ARTs) or be cryopreserved in biobanks for their future use. Traditionally, the minimally invasive technique of choice performed in vivo in domestic and wild felines was LOPU (laparoscopic-guided ovum pick up). The present study was designed to explore if ultrasound-guided OPU in the domestic cat is safe and effective. In an initial series of ex vivo experiments (n = 92 ovaries, n = 434 oocytes), the effect of different aspiration pressures for oocyte collection was explored. These experiments identified 43 mmHg as the optimal aspiration pressure, resulting in the highest recovery rate and a favorable maturation and blastocyst rate. Subsequently, 16 grade I and II oocytes were retrieved by OPU and 101 oocytes were retrieved following ovariectomy and slicing. Sixteen oocytes obtained with each technique were subjected to in vitro maturation (IVM) and in vitro fertilization (IVF). A total of 14 presumptive zygotes were selected for in vitro culture (IVC) from each group (OPU and slicing), obtaining a cleavage rate of 57.1 % and 64.2 %, a morula rate of 28.5 % in both groups, and a blastocyst rate of 7.14 % and 14.2 % respectively. The hormonal stimulation protocol was well-tolerated, with no adverse effects observed. Moreover, no complications arose during the ovariectomy performed post-OPU. The use of this technique in domestic cats represents a significant step forward in terms of safety, replicability, and invasiveness, serving as a valuable model for its application in wild felids species. Additional research involving a greater number of animals is required to validate these encouraging findings.
卵母细胞采集（OPU）是一种广泛应用于牛和母马的微创技术，用于获取卵母细胞，包括在超声引导下对卵巢卵泡进行穿刺。已经证明，这种技术可以安全地在同一头雌性动物身上重复使用，而不会影响其生殖健康，可以在不考虑其生殖状态的情况下从个体中获取卵母细胞。通过 OPU 获得的卵母细胞随后可以用于体外胚胎生产（IVP），使用辅助生殖技术（ART），或者在生物库中冷冻保存以备将来使用。传统上，在国内和野生猫科动物中进行的微创技术选择是 LOPU（腹腔镜引导的卵母细胞采集）。本研究旨在探讨在国内猫中进行超声引导的 OPU 是否安全有效。在一系列初步的离体实验中（n=92 个卵巢，n=434 个卵母细胞），研究了不同的抽吸压力对卵母细胞采集的影响。这些实验确定 43mmHg 为最佳抽吸压力，可获得最高的回收率以及有利的成熟和囊胚率。随后，通过 OPU 采集了 16 个 I 级和 II 级卵母细胞，并通过卵巢切除术和切片采集了 101 个卵母细胞。用每种技术获得的 16 个卵母细胞进行体外成熟（IVM）和体外受精（IVF）。从每组（OPU 和切片）中选择 14 个推定受精卵进行体外培养（IVC），分别获得 57.1%和 64.2%的卵裂率、28.5%的桑椹胚率和 7.14%和 14.2%的囊胚率。激素刺激方案耐受性良好，未观察到不良反应。此外，OPU 后进行的卵巢切除术中未出现并发症。该技术在国内猫中的应用在安全性、可重复性和微创性方面是一个重大进展，为其在野生猫科物种中的应用提供了一个有价值的模型。需要更多的动物研究来验证这些令人鼓舞的发现。
