Diagnostic epididymal and testicular sperm recovery and genetic aspects in azoospermic men

University of Gothenburg, Goeteborg, Västra Götaland, Sweden
Human Reproduction (Impact Factor: 4.57). 02/1999; 14(1). DOI: 10.1093/humrep/14.1.118
Source: OAI


Various procedures for sperm recovery in azoospermic men have been described, from open testicular biopsy to simple needle aspiration from the epididymis and the testis. Fifty-one obstructive and 86 non-obstructive azoospermic men were treated to compare the recovery of spermatozoa obtained by percutaneous aspiration from the epididymis (PESA) and aspiration/extraction from the testis (TESA, TESE) with histopathology. If TESA failed, the work up proceeded with TESE. All patients were karyotyped. Spermatozoa were recovered by PESA or TESA in all obstructive men (51/51 patients). In 22 out of 86 patients with non-obstructive azoospermia, testicular spermatozoa could be successfully recovered by TESA. In five additional patients TESE was successful in recovering spermatozoa where TESA had failed. In 43 patients, neither TESA nor TESE was successful. Sixteen patients chose not to proceed with TESE. Seven out of 86 patients had an abnormal karyotype in the non-obstructive group (8%), none in the obstructive group. In the non-obstructive patient group testicular histopathology showed hypospermatogenesis, incomplete maturation arrest and germ cell aplasia with focal spermatogenesis in cases where spermatozoa were recovered and complete germ cell aplasia, complete maturation arrest and fibrosis in cases where no spermatozoa were found. Spermatozoa were recovered by PESA or TESA from all patients with obstructive azoospermia and from ~40% of patients with non-obstructive azoospermia by TESA or TESE. Retrieval of viable spermatozoa in the infertility work-up was highly predictable for sperm recovery in subsequent ICSI cycles. TESA performed under local anaesthesia seems almost as effective as more invasive procedures in recovering testicular spermatozoa, both in obstructive and non-obstructive azoospermic men.

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    • "Of all infertile marriages, estimated at 10–15 % of total, about half include male factor etiology (Hull et al., 1985; ESHRE Capri Workshop, 1996). In about 60 % of azoospermic males, an obvious clinical reason, such as an obstruction in the efferent ducts, cannot be found (Westlander et al., 1999). For this group, the diagnosis idiopathic male infertility is coined, unless a genetic factor such as deletions of the azoospermia factors (AZF) on the Y chromosome (15–20 % in azoospermic probands and 7–10 % in severe oligozoospermia, Krausz et al., 2000) or an abnormal karyotype (4–12 %, de Braekeleer and Dao, 1991; Tuerlings et al., 1998) can be identified. "
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    ABSTRACT: We have developed a protocol for the identification of aberrant chromosome behavior during human male meiosis up to metaphase of the secondary spermatocyte. Histological evaluation by the Johnsen score of a testicular biopsy was combined with immunofluorescence of first meiotic prophase spermatocytes, using antibodies against synaptonemal complex protein 3 (SYCP3) and the product of the ataxia telangiectasia and rad3-related gene (ATR). This combination enables accurate meiotic prophase substaging and the identification of pachytene spermatocytes with asynapsis. Furthermore, we also investigated the competence of late pachytene primary spermatocytes to complete the first meiotic division up to metaphase and of secondary spermatocytes to transform into metaphase by an in vitro challenge with okadaic acid (OA). We tested this protocol on five males with normal Johnsen scores that presented with obstructive azoospermia, five males with low Johnsen scores and non-obstructive azoospermia and six vasectomized control males of proven fertility and normal Johnsen scores. In all azoospermics, the profiling of meiotic prophase stages by immunofluorescence increases the resolving power of the Johnsen score. In both obstructive and non-obstructive azoospermic patients, relatively more leptotene meiotic prophase stages were counted compared to the controls. In non-obstructive azoospermics, a marked heterogeneity in spermatogenesis was found, after combining the results of all three approaches, pointing at functional mosaicism of the germinal epithelium. Asynaptic pachytene spermatocytes were rarely encountered. Also, when first meiotic metaphase could be induced by OA, chiasma counts were normal. In none of the non-obstructive azoospermic males did the pattern of spermatogenesis resemble that of knock-out mouse azoospermics. We conclude that this combined histological and cytological approach enables a detailed phenotypic classification of infertile males, at a level comparable to that applied for male-sterile knock-out mice with a meiotic defect. This may facilitate the identification of candidate genes for human male infertility.
    Full-text · Article · Feb 2004 · Cytogenetic and Genome Research
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    • "In cases of obstructive azoospermia (OA), sufficient numbers of spermatozoa for ICSI can usually be retrieved by epididymal sperm aspiration, whereas cases of NOA mostly require open testicular biopsy. Even with this more invasive approach, spermatozoa fail to be recovered in about a half of the cases of NOA in tissue samples obtained by multiple-site testicular biopsy (Kahraman et al, 1996; Westlander et al, 1999; Friedler et al, 2002). Unfortunately, a reliable test predicting successful sperm retrieval from the testes of men with NOA is still lacking. "

    Preview · Article · May 2003 · Journal of Andrology
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    • "Currently, there are no clinical or laboratory methods that can reliably and accurately predict the presence of sperm on testicular sperm extraction (TESE). In up to 60% of the patients suffering from non-obstructive azoospermia (NOA), no mature sperm are found that would enable ICSI to be attempted, despite several biopsies taken (Devroey et al., 1995; Kahraman et al., 1996; Friedler et al., 1997; Schlegel et al., 1997; Silber et al., 1997; Tournaye et al., 1997; Rosenlund et al., 1998; Westlander et al., 1999). Whether a repetitive TESE should be recommended in patients that had no mature sperm following their first TESE remains an unanswered question. "
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    ABSTRACT: BACKGROUND: It is unclear whether or not testicular sperm extraction (TESE) should be repeated for patients in whom no sperm were found during their first TESE attempt. METHODS AND RESULTS: The outcome of repeated TESE was evaluated in patients with non-obstructive azoospermia (NOA) after failing to obtain sperm in their first extraction attempt, or having used all available cryopreserved testicular tissue. Out of 83 patients with NOA, patients repeated TESE two (n = 22), three (n = 8), four (n = 6) and five (n = 3) times. Distribution of main testicular histology included germ cell aplasia (55%), maturation arrest (29%) and germ cell hypoplasia (16%). The first TESE yielded mature sperm for ICSI in 39% of patients (sp+), and failed in the remaining 61% (sp–). A second TESE yielded mature sperm in 1/4 from the sp– group and in 16/18 from the sp+ group. At the third, fourth and fifth trials, 8/8, 5/6 and 3/3 of the original sp+ patients were sp+ again respectively. Compared with the outcome of the first trial, all further trials did not differ statistically in the rate of fertilization (54 versus 49%), implantation (9.5 versus 5.4%), or clinical pregnancy/cycle (19 versus 15%). No pregnancies were achieved among the three patients after their fifth TESE. Pregnancies occurred in all histological groups, except maturation arrest. CONCLUSIONS: The outcome of repeated TESE cycles, up to the fourth trial, justifies the procedure.
    Full-text · Article · Sep 2002 · Human Reproduction
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