Quantification of seminal germ cells in azoospermia: correlations with testicular histology and TESE outcome

Abstract

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.

Molecular analysis of testis biopsy and semen pellet as complementary methods with histopathological analysis of testis in non-obstructive azoospermia

“…Yeung et al showed that quantification of seminal IGCs in azoospermia using flow cytometry cannot predict TESE outcome. According to their conclusion, this inability could be due to the limitations of flow cytometry in differentiating between germ cells and other artifacts in semen precipitates [26]. In our pervious study, we demonstrated that concomitant detection of DAZ and PRM2 transcripts by RT-PCR in seminal fluid is able to predict the presence of spermatid/sperm in testis of NOA men [18].…”

Cytogenetic and molecular analyses of de novo translocation dic(9;13)(p11.2;p12) in an infertile male

“…The probability rate for unbalanced progeny with trisomy/monosomy of the 9p11.2 segment after adjacent-1 segregation at birth for paternal carriers was estimated at 11.8%, and the odds of unbalanced progeny at prenatal diagnosis was 57% [49]. The probability of natural conception was very low, but an option for ICSI after testicular biopsy and pre-implantation genetic diagnosis was discussed with the couple because the presence of immature germ cells in the ejaculate could indicate spermatogenic activity in the testes [50]. Unfortunately, bilateral testicular biopsies did not reveal any sperm cells (N. Huleyuk, personal communication).…”

Prognostic markers for competent human spermatozoa: fertilizing capacity and contribution to the embryo

“…The latter score count reveals close correlation with TESE results when using cryopreserved testicular tissue for diagnostic purposes before hormonal stimulation of the female partner and ICSI (unpublished data). Detection of immature germ cells in semen, however, does not provide a prognostic marker for successful TESE, although their presence could indicate the spermatogenic activity within the testis (Yeung et al. , 2007).…”