Yeung C H, Beiglböck-Karau L, Luetjens C M, Wunsch A, Nieschlag E
Institute of Reproductive Medicine, University of Münster, Münster, Germany.
Int J Androl. 2009 Jun;32(3):242-54. doi: 10.1111/j.1365-2605.2007.00843.x. Epub 2007 Dec 7.
In the treatment of male infertility by intra-cytoplasmic injection of spermatozoa (ICSI) extracted from testicular tissue (TESE), the high incidence of negative TESE outcome calls for non-invasive prognostic methods. Literature suggests that seminal haploid germ cell detection could be one. For this purpose, a multi-parametric stringent flow cytometric method was applied to 50 TESE patients for the quantification of ejaculated germ cells. Cells from 50 ejaculates were identified and quantified as spermatozoa (HC, highly condensed), round spermatids (1N), primary spermatocytes (SPC) (4N) or diploid cells (2N, including somatic and non-testicular cells) by their DNA and mitochondria staining and laser scatter characteristics, and compared with testicular biopsy histology and TESE outcome. Whereas 96% of patients displayed a diploid peak in the distribution histograms, the HC, 1N and 4N peaks were absent from the majority of samples. In 13 ejaculates, either a HC or 1N or 4N peak, or a combination of these, was discernible. Although seminal germ cell numbers bore no overall association with elongated spermatids (ES) in histology or spermatozoa retrieval in TESE outcome, 4N cells per ejaculate were correlated with the percentage of tubule sections showing SPC as the most advanced germ cells. The incidence of HC peaks was higher in patients showing some ES in histology or sperm retrieval than in the sperm-negative groups. In groups with suspected obstruction showing nearly full spermatogenesis and maximal sperm retrieval, there was no incidence of a HC peak. Germ cell peaks were associated with germ cell degeneration noted in testicular histology. In conclusion, seminal germ cells cannot provide good prognosis for TESE, although their presence could indicate the spermatogenic activity in the testis.
在通过胞浆内注射从睾丸组织中提取的精子（TESE）治疗男性不育症时，TESE阴性结果的高发生率需要非侵入性的预后方法。文献表明，精液单倍体生殖细胞检测可能是一种方法。为此，将一种多参数严格流式细胞术方法应用于50例TESE患者，以定量射精的生殖细胞。通过DNA和线粒体染色以及激光散射特征，对50份射精样本中的细胞进行鉴定和定量，确定为精子（HC，高度浓缩）、圆形精子细胞（1N）、初级精母细胞（SPC）（4N）或二倍体细胞（2N，包括体细胞和非睾丸细胞），并与睾丸活检组织学和TESE结果进行比较。虽然96%的患者在分布直方图中显示出二倍体峰，但大多数样本中没有HC、1N和4N峰。在13份射精样本中，可以辨别出HC峰、1N峰或4N峰，或这些峰的组合。虽然精液生殖细胞数量与组织学中的细长精子细胞（ES）或TESE结果中的精子获取总体上没有关联，但每份射精中的4N细胞与显示SPC为最成熟生殖细胞的小管切片百分比相关。在组织学或精子获取中显示有一些ES的患者中，HC峰的发生率高于精子阴性组。在疑似梗阻且显示几乎完全精子发生和最大精子获取的组中，没有HC峰的发生。生殖细胞峰与睾丸组织学中观察到的生殖细胞退化有关。总之，精液生殖细胞不能为TESE提供良好的预后，尽管它们的存在可能表明睾丸中的生精活性。
