Esbert M, Godo A, Soares S R, Florensa M, Amorós D, Ballesteros A, Vidal F
IVI Barcelona, Barcelona, Spain.
Genetics of Male Fertility Group, Unitat de Biologia Cellular (Facultat de Biociències), Universitat Autònoma de Barcelona, Bellaterra, Spain.
Andrology. 2017 Jul;5(4):807-813. doi: 10.1111/andr.12376. Epub 2017 Jun 14.
Colloidal super-paramagnetic microbeads conjugated with annexin V are effective for separating apoptotic spermatozoa by MACS as a result of the high affinity of annexin V for externalized PS molecules. The effectiveness of the procedure in reducing the percentage of sperm with fragmented DNA and abnormal morphology has also been reported. However, it is still unknown if it could decrease the percentage of aneuploid spermatozoa. The objective of our prospective study, performed on 16 males with abnormal FISH on spermatozoa, was to assess if MACS columns were useful tools to retain spermatozoa carrying chromosomal abnormalities in semen samples processed after density gradient centrifugation (DGC). The pellet obtained after DGC was subjected to MACS, and sperm FISH analyses were performed both in the eluded fraction and in the fraction retained in the column. The observed frequencies of disomy and nullisomy 13, 18, and 21, X and Y, as well as the diploidy rates in the MACS eluded fraction and the fraction retained in the MACS column were recorded. We observed that the frequencies of aneuploidies in the eluded fraction were lower than in the fraction retained in the MACS column (0.59% vs. 0.75%; p = 0.010). DGC determined a significant reduction in sperm concentration (z-ratio = 2.83; p = 0.005) and a significant increase in sperm progressive motility (z-ratio = -3.5; p < 0.001). MACS also led to a significant reduction in sperm concentration (z-ratio = 3.14; p = 0.002) and a significant increase in progressive motility (z-ratio = -2.59; p = 0.01) when compared with the post-DGC sample. Sperm concentration was similar in the two fractions generated by MACS (z-ratio = 0.63; p = 0.52), while progressive motility was significantly higher in the MACS eluded fraction (z-ratio = 2.42; p = 0.02). According to our results, MACS columns are able to selectively retain spermatozoa carrying chromosomal abnormalities. Furthermore, the performance of DGC and MACS on semen samples leads to an enrichment of progressive motility.
由于膜联蛋白V对外化的磷脂酰丝氨酸（PS）分子具有高亲和力，与膜联蛋白V结合的胶体超顺磁性微珠可通过磁性激活细胞分选（MACS）有效地分离凋亡精子。该方法在降低DNA片段化和形态异常精子百分比方面的有效性也已有报道。然而，它是否能降低非整倍体精子的百分比仍不清楚。我们对16名精子荧光原位杂交（FISH）异常的男性进行了前瞻性研究，目的是评估MACS柱是否是在密度梯度离心（DGC）后处理的精液样本中保留携带染色体异常精子的有用工具。DGC后获得的沉淀进行MACS处理，并对洗脱部分和保留在柱中的部分进行精子FISH分析。记录在MACS洗脱部分和保留在MACS柱中的部分中观察到的13、18和21号染色体、X和Y染色体的二体性和单体性频率，以及二倍体率。我们观察到，洗脱部分的非整倍体频率低于保留在MACS柱中的部分（0.59%对0.75%；p = 0.010）。DGC导致精子浓度显著降低（z比值 = 2.83；p = 0.005），精子前向运动能力显著增加（z比值 = -3.5；p < 0.001）。与DGC后的样本相比，MACS也导致精子浓度显著降低（z比值 = 3.14；p = 0.002），前向运动能力显著增加（z比值 = -2.59；p = 0.01）。MACS产生的两个部分中的精子浓度相似（z比值 = 0.63；p = 0.52），而MACS洗脱部分的前向运动能力显著更高（z比值 = 2.42；p = 0.02）。根据我们的结果，MACS柱能够选择性地保留携带染色体异常的精子。此外，DGC和MACS对精液样本的处理导致前向运动能力的富集。
