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单操作者经皮卵巢穿刺技术中抽吸压力、卵泡冲洗方法及针头旋转对母马卵母细胞回收和胚胎生产的影响。

The Influence of Aspiration Pressure, Follicle Flushing Method and Needle Rotation During Single-Operator OPU Technique on Oocyte Recovery and Embryo Production in the Mare.

作者信息

Cuervo-Arango Juan, Sala-Ayala Laura, Márquez-Moya Adrián, Martínez-Boví Rebeca

机构信息

Equine Fertility Group, Faculty of Veterinary Medicine, Universidad Cardenal Herrera-CEU, CEU Universities, 46115 Alfara del Patriarca, Valencia, Spain.

出版信息

Animals (Basel). 2025 Mar 14;15(6):832. doi: 10.3390/ani15060832.

Abstract

The objective of this study was to determine the effect of two aspiration pressures (75 vs. 150 mmHg), the follicle flushing method (injection pump controlled by a foot pedal vs. a plastic syringe) and the twisting of the OPU needle on oocyte recovery and in vitro embryo production. OPU data from a total of 104 warmblood sport mares belonging to a commercial OPU-ICSI program were collected as part of a prospective study split into three experiments. Each mare was used only once for OPU. In Experiment 1, the mares' follicles were aspirated using either a high aspiration pressure (flow rate of 1.33 mL/s; = 18) or low aspiration pressure (0.75 mL/s; = 18); in Experiment 2, follicles were flushed using either a manual method (plastic syringe, = 18) or an automatic method (injection pump controlled by a foot pedal, = 18); and in Experiment 3, the follicles were aspirated by scraping the follicle wall with needle rotation (needle twisting, = 16) or without needle rotation (control, = 16). In all the experiments, the same OPU operator and technician searching oocytes were used, and the allocation of each mare to the different treatment groups was randomized. The overall mean oocyte recovery rate of the study was 54.2 ± 17.1%, and the mean number of embryos per OPU-ICSI session was 1.9 ± 1.6. The oocyte recovery rate was not influenced by any of the parameters investigated ( > 0.05). However, high aspiration pressure (150 mmHg) tended to yield oocytes with lower maturation (51.6%; = 0.09) and blastocyst rates (20.6%; = 0.08) following IVM and ICSI, respectively, compared with the low aspiration group (64.4% MII rate and 31.4% blastocyst rate). In conclusion, increasing aspiration pressure does not increase oocyte recovery. Furthermore, when a single operator performs the OPU (holding the ovary and handling the needle simultaneously), needle rotation to scrape the follicle wall does not improve oocyte recovery.

摘要

本研究的目的是确定两种抽吸压力(75 mmHg与150 mmHg)、卵泡冲洗方法(脚踏控制的注射泵与塑料注射器)以及经皮卵巢穿刺取卵(OPU)针的扭转对卵母细胞回收及体外胚胎生产的影响。作为一项前瞻性研究的一部分,收集了总共104匹属于商业OPU-卵胞浆内单精子注射(ICSI)项目的温血运动母马的OPU数据,该研究分为三个实验。每匹母马仅用于一次OPU。在实验1中,使用高抽吸压力(流速1.33 mL/s;n = 18)或低抽吸压力(0.75 mL/s;n = 18)抽吸母马的卵泡;在实验2中,使用手动方法(塑料注射器,n = 18)或自动方法(脚踏控制的注射泵,n = 18)冲洗卵泡;在实验3中,通过旋转针头刮擦卵泡壁(针头扭转,n = 16)或不旋转针头(对照,n = 16)抽吸卵泡。在所有实验中,使用相同的OPU操作员和寻找卵母细胞的技术人员,并且每匹母马分配到不同治疗组是随机的。该研究的总体平均卵母细胞回收率为54.2±17.1%,每次OPU-ICSI操作的平均胚胎数为1.9±1.6。卵母细胞回收率不受所研究的任何参数影响(P>0.05)。然而,与低抽吸组相比,高抽吸压力(150 mmHg)在体外成熟(IVM)和ICSI后分别倾向于产生成熟率较低(51.6%;P = 0.09)和囊胚率较低(20.6%;P = 0.08)的卵母细胞(低抽吸组的MII率为64.4%,囊胚率为31.4%)。总之,增加抽吸压力不会提高卵母细胞回收率。此外,当单一操作员进行OPU(同时握住卵巢并操作针头)时,旋转针头刮擦卵泡壁并不能提高卵母细胞回收率。

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