Fertilis Pty Ltd, Frome Road, Helen Mayo South, The University of Adelaide, Adelaide, SA, 5005, Australia.
Virtual Ark Pty Ltd, 73 Woolnough Road, Semaphore, SA, 5019, Australia.
J Assist Reprod Genet. 2024 Feb;41(2):297-309. doi: 10.1007/s10815-023-03018-0. Epub 2024 Jan 18.
Intracytoplasmic sperm injection (ICSI) imparts physical stress on the oolemma of the oocyte and remains among the most technically demanding skills to master, with success rates related to experience and expertise. ICSI is also time-consuming and requires workflow management in the laboratory. This study presents a device designed to reduce the pressure on the oocyte during injection and investigates if this improves embryo development in a porcine model. The impact of this device on laboratory workflow was also assessed.
Porcine oocytes were matured in vitro and injected with porcine sperm by conventional ICSI (C-ICSI) or with microICSI, an ICSI dish that supports up to 20 oocytes housed individually in microwells created through microfabrication. Data collected included set-up time, time to align the polar body, time to perform the injection, the number of hand adjustments between controllers, and degree of invagination at injection. Developmental parameters measured included cleavage and day 6 blastocyst rates. Blastocysts were differentially stained to assess cell numbers of the inner cell mass and trophectoderm. A pilot study with human donated MII oocytes injected with beads was also performed.
A significant increase in porcine blastocyst rate for microICSI compared to C-ICSI was observed, while cleavage rates and blastocyst cell numbers were comparable between treatments. Procedural efficiency of microinjection was significantly improved with microICSI compared to C-ICSI in both species.
The microICSI device demonstrated significant developmental and procedural benefits for porcine ICSI. A pilot study suggests human ICSI should benefit equally.
胞质内精子注射(ICSI)会对卵母细胞的卵母细胞膜造成物理压力,并且仍然是最具挑战性的技术之一,其成功率与经验和专业知识有关。ICSI 也很耗时,并且需要在实验室中进行工作流程管理。本研究介绍了一种旨在减少注射过程中对卵母细胞压力的装置,并研究了它是否能改善猪模型中的胚胎发育。还评估了该装置对实验室工作流程的影响。
猪卵母细胞在体外成熟,并通过常规 ICSI(C-ICSI)或微 ICSI 进行注射,微 ICSI 是一种支持多达 20 个卵母细胞单独放置在微井中的 ICSI 培养皿,这些微井是通过微加工创建的。收集的数据包括设置时间、对准极体的时间、进行注射的时间、控制器之间的手动调整次数以及注射时的凹陷程度。测量的发育参数包括卵裂和第 6 天囊胚率。囊胚通过差异染色来评估内细胞团和滋养外胚层的细胞数量。还对用人捐赠的 MII 卵母细胞注射珠子进行了初步研究。
与 C-ICSI 相比,微 ICSI 显著提高了猪囊胚率,而卵裂率和囊胚细胞数量在两种处理方式之间无差异。与 C-ICSI 相比,微 ICSI 显著提高了微注射的程序效率。
微 ICSI 装置为猪 ICSI 提供了显著的发育和程序优势。初步研究表明,人类 ICSI 也应同样受益。