Department of Anatomy, Iran University of Medical Science, Tehran, Iran.
Endometrium & Endometriosis Research Center, Hamadan University of Medical Sciences, Hamadan, Iran.
Hum Fertil (Camb). 2023 Dec;26(6):1408-1416. doi: 10.1080/14647273.2023.2236297. Epub 2023 Jul 20.
Conventional sperm selection based on motility and morphology fails to provide detailed information on sperm functional and molecular status. Magnetic-activated cell sorting (MACS) protocol aims to optimize this process by selecting apoptotic sperm cells. Phospholipase C zeta-1 (PLCz1) is a physiological stimulus for oocyte activation and early embryonic development. The purpose of this study was to examine seminal parameters, DNA fragmentation index (DFI), and PLCz1 expression levels in MACS-DGC sorted specimens (DFI > 30%) and assess early development in resulting embryos. Semen specimens from 60 patients diagnosed with male factor infertility were collected and processed by either density gradient centrifugation (DGC) or MACS-DGC protocols. Pre and post-preparation analysis was performed. PLCz1 expression was assessed using the RT-PCR method. Retrieved eggs from their partners were divided into two groups in which they were injected with different sorted sperm. The fertilization rate and embryonic development were evaluated. While sperm's progressive motility and morphology significantly improved, there was a substantial decline in DFI following MACS-DGC. Fertilization rates were almost the same between the groups, and the latter resulted in remarkably more top-quality embryos and more blastocysts. PLCz1 expression was considerably higher in the MACS-DGC group. By eliminating apoptotic cells, the MACS-DGC technique could sort highly PLCz1-expressed sperm, optimize sperm selection in individuals with elevated DFI, development of resulting embryos.
基于活力和形态的传统精子选择无法提供精子功能和分子状态的详细信息。磁激活细胞分选 (MACS) 方案旨在通过选择凋亡精子细胞来优化此过程。磷酯酶 C ζ-1 (PLCz1) 是卵母细胞激活和早期胚胎发育的生理刺激物。本研究的目的是检查 MACS-DGC 分选标本中的精液参数、DNA 碎片化指数 (DFI) 和 PLCz1 表达水平(DFI>30%),并评估由此产生的胚胎的早期发育。收集了 60 名被诊断为男性因素不育症的患者的精液标本,并通过密度梯度离心 (DGC) 或 MACS-DGC 方案进行处理。进行了预处理和预处理后的分析。使用 RT-PCR 方法评估 PLCz1 表达。从其伴侣中取出的卵子分为两组,用不同的分选精子进行注射。评估受精率和胚胎发育情况。虽然精子的前向运动和形态显著改善,但 MACS-DGC 后 DFI 大幅下降。两组的受精率几乎相同,但后者产生了更多的优质胚胎和更多的囊胚。MACS-DGC 组中 PLCz1 的表达明显更高。通过消除凋亡细胞,MACS-DGC 技术可以分选高 PLCz1 表达的精子,优化 DFI 升高的个体中的精子选择,从而优化由此产生的胚胎的发育。