PURPOSE: We determined the usefulness of in vitro germ cell culture in nonobstructive azoospermic patients diagnosed with Sertoli cell only syndrome, no sperm in testicular sperm extraction. MATERIALS AND METHODS: This study included 44 patients (45 testicular tissues) with nonobstructive azoospermia who were diagnosed with Sertoli cell only syndrome and were found to have no sperm in testicular sperm extraction between January 2006 and July 2008. Among the 45 testicular tissues, 22 tissues were processed for culture. In the in vitro cultures, the testicular tissues were dissociated and plated on gelatin-coated dishes. Patients were divided into 2 groups according to culture success: group I, culture positive (+; n=10); and group II, culture negative (-; n=12). RESULTS: The mean patient ages were 31.73 and 31.68 years for groups I and II, respectively. The mean testicular sizes were 10.19 and 10.42 cc, respectively; the semen volumes were 2.86 and 3.04 cc, respectively; and the mean FSH, LH, and testosterone levels were 18.86 mIU/ml, 5.99 mIU/ml, and 4.46 ng/ml vs. 21.02 mIU/ml, 6.29 mIU/ml, and 4.32 ng/ml for groups I and II, respectively, with no significant differences between the groups (p>0.05). The culture rate of nonobstructive azoospermic patients diagnosed with Sertoli cell only syndrome was 45.5% (10/22). Round spermatid injection was done in 2 patients with consent of the patients, but implantation failed. Among the 45 tissues, germ cells were found in 8 tissues after pathologic reexamination. CONCLUSIONS: The in vitro culture of germ cells would be useful in the advanced treatment of nonobstructive azoospermic patients.
Azoospermia ; Germ Cells ; Humans ; Semen ; Sertoli Cell-Only Syndrome ; Spermatids ; Spermatozoa ; Testosterone

PURPOSE: Because testicular biopsy traditionally has been performed as a separate operation, there are some needs for immediate interpretation of the testicular biopsy specimen in obstructive azoospermia patients. The realization that a proportion of patients with nonobstructive azoospermia harbor spermatozoa in their testicular parenchyma, combined with the ability of intracytoplasmic sperm injection(ICSI) to effect pregnancy with single sperm, has prompted clinicians to explore testicular sperm extraction(TESE) in these patients. We performed this study to investigate the meaning of touch imprint and TESE. MATERIALS AND METHODS: Fifty-two patients with azoospermia underwent touch imprint, TESE and simultaneous formal testicular biopsy. Testicular biopsy tissue was touched on a slide and stained using a Diff-Quik method. Tissue obtained from TESE was analyzed by intensive searching to find spermatozoa. RESULTS: Twenty-nine patients were determined as obstructive azoospermia including two cases of hypospermatogenesis, all patients had sperm seen on touch imprint and TESE. The other patients were diagnosed as nonobstructive azoospermia, testicular histology was divided into two main patterns: Sertoli cell only syndrome, spermatocytic arrest. Touch imprint failed to show sperm, but three of twenty patients(15%) with Sertoli cell only syndrome had sperm present during TESE. CONCLUSIONS: Touch imprint provides high quality cellular detail for immediate review, which can minimize the intraoperative time required to decide upon the appropriateness of correcting the ductal obstruction in obstructive azoospermia patients. Men with nonobstructive azoospermia may have mature spermatozoa present within their testicular parenchyma. Therefore intensive searching for presence of spermatozoa should be performed in tissue obtained from TESE by trained specialist.
Azoospermia* ; Biopsy* ; Humans ; Male ; Oligospermia ; Pregnancy ; Sertoli Cell-Only Syndrome ; Specialization ; Spermatozoa* ; Testis*

There were 122 infertile males who received testicular biopsy in our department during the period from 1978 to 1980. Among them, 20 patients were diagnosed to have Sertoli cell only syndrome by pathologic finding. A cumulative clinical investigation was undertaken in 20 patients with Sertoli cell only syndrome using history taking, orchidometer, semen analysis, and plasma hormonal assay (FSH. LH, & Testosterone). We found the interesting relationship between the size of the testis and plasma hormonal levels. 1. In pathologic finding, Sertoli cell only syndrome was 16%, spermatic arrest 16%, hypospermatogenesis 15%, peritubular fibrosis 15%, and normal testis accompanying obstruction of efferent duct 22%. 2. The size of the testis was average 12ml, and plasma FSH was increased to average 27.43 IU/L, plasma LH was also increased to average 25.9 IU/L, but plasma testosterone was within normal ranges (average 5.58 ng/ml).
Biopsy ; Fibrosis ; Humans ; Male ; Oligospermia ; Plasma* ; Reference Values ; Semen Analysis ; Sertoli Cell-Only Syndrome* ; Testis* ; Testosterone

Investigations into the pathogenesis of male infertility have relied predominantly on two types of analysis: histological evaluations of testicular biopsies and assays of these hormones thought to be responsible for the regulation of spermatogenesis. Other types of analysis such as chromosomal studies and biochemical determinations of ejaculate components have also played a role in characterizing male infertility. Testis biopsy findings of 54 cases with azoospermia were observed in aspect of testicular size and pastory and seminovesiculograms were performed in 3 cases with normal testis and 1 case with peritubular fibrosis and reduced spermatogenesis. The following results were obtained. 1) Among the 42 cases with normal sized testis, 12 cases (28.6%) of normal testis, 10 cases (23.8%) of peritubular fibrosis with reduced spermatogenesis, 11 cases (26. 2%) of reduced spermatogenesis, 1 case(2.4%) of maturation arrest at spermatid level, 6 cases (14.3%) of Sertoli cell only syndrome and 2cases (4.7%) of hyalinized tubule with Leydig cell hyperplasia were observed. 2) Among the 12 cases with small sized testis, 1 case (8.3%) of normal testis, 1 case (8.3%) of peritubular fibrosis with reduced spermatogenesis, 2 cases(16.7%) of reduced spermatogenesis, 5 cases(41.7%) of Sertoli cell only syndrome and 3 cases (25.O%) of hyalinized tubule with Leydig cell hyperplasia were observed. 3) Normal testis was observed in 4 of 9 cases with tuberculous epididymitis and 1 case with surgically corrected bilateral hydrocele on their past history. Peritubular fibrosis with reduced spermatogenesis was observed in 3 of 9 cases with urethritis, 2of3 cases with tuberculous epididymitis and 1 case with surgically corracted bilateral varicocele on their past history. 4) The observed in 3 cases of normal testis and 1 carmel testis and 1 case of peritubular fibrosis with reduced spermatogenesis on the seminovesiculogram.
Azoospermia* ; Biopsy* ; Epididymitis ; Fibrosis ; Humans ; Hyalin ; Hyperplasia ; Infertility ; Infertility, Male ; Male ; Sertoli Cell-Only Syndrome ; Spermatids ; Spermatogenesis ; Testis* ; Urethritis ; Varicocele

ObjectiveTo investigate the 5'-flanking regulatory sequence methylation status of the Boule gene in the testis tissue of infertile men with Sertoli cell-only syndrome (SCOS).

METHODSWe collected biopsy samples of the testis tissue from 12 men with obstructive azoospermia (the control group) and 15 cases of SCOS, all without varicocele, cryptorchidism, or infectious disease. We extracted genomic DNA from the testis tissue of the SCOS patients, analyzed the characteristics of the 5'-flanking regulatory sequence of the Boule gene using the bioinformatics method, and detected the methylation status of the Boule gene by sodium bisulfite sequencing.

RESULTSA CpG island was observed in the 5'-flanking regulation region of the Boule gene. The methylation level of the Boule gene was remarkably higher in the SCOS group than in the obstructive azoospermia controls (61.4% vs 21.7%, P<0.01), with significant differences in the methylation levels of 14 CpG sites, namely, －58 bp, －50 bp, －48 bp, －38 bp, －28 bp, －24 bp, －20 bp, －15 bp, －1 bp, +5 bp, +8 bp, +15 bp, +29 bp, and +58 bp.

CONCLUSIONSThe methylation level of the Boule gene is significantly higher in the SCOS patients than in the obstructive azoospermia males, which suggests that the changes in Boule methylation may be associated with spermatogenic dysfunction.

Case-Control Studies ; DNA Methylation ; Humans ; Male ; RNA-Binding Proteins ; genetics ; Sertoli Cell-Only Syndrome ; genetics ; Spermatogenesis ; Testis ; metabolism

This study was carried to determine the possibility of finding motile spermatozoa and fertilization, pregnancy rate after testicular sperm extraction(TESE) with ICSI in obstructive and non-obstructive azoospermic patients. In 154 cases(132 patients), obstructive azoospermia was 77 cases and non-obstructive azoospermia was 77 cases. In obstructive azoospermia, patients generally showed normal spermatogenesis and included vas agenesis(n=8), multiple vas obstruction(n=7), epididymal obstruction (n=54). Total of 982 retrieved oocytes were obtained and 84.4% were injected. The fertilization rates with 2 PN and cleavage rate were 72.5% and 62.3%, .respectively. 30 pregnancies(38.9%) were achieved and the ongoing pregnancies were 22 cases (28.6%). In non-obstructive azoospermia, patients showed hypospermatogenesis(n=49), maturation arrest(n=4), Sertoli cell only syndrome (n=24). The various stages of spermatogenic cell could be retrieved by TESE and could be reached normal fertilization and embryo development with ICSI. Total of 1072 retrieved oocytes obtained and 80.2% were injected. The fertilization rates with 2 PN and cleavage rate were 52.8% and 68.9%, respectively. 22 pregnancies(30.1%) were achieved and the ongoing pregnancies were 19 cases(26.0%). Conclusively, the combination of TESE with ICSI using testicular spermatozoa can achieve normal fertilization and pregnancy rate and effective method in obstructive and non-obstructive azoospermic patients.
Azoospermia ; Embryonic Development ; Female ; Fertilization* ; Humans ; Oocytes ; Pregnancy Rate* ; Pregnancy* ; Sertoli Cell-Only Syndrome ; Sperm Injections, Intracytoplasmic ; Spermatogenesis ; Spermatozoa*

Irreparable obstructive azoospermic patients can be treated successfully with microsurgical epididymal sperm aspiration(MESA) o. testicular sperm extraction (TESE) by intracytoplasmic sperm injection(ICSI). Obstructive azoospermic patients generally have normal spermatogenesis. The aim of this study was to see if any spermatozoa could be retrieved from non-obstructive azoospermia and to assess the efficacy of ICSI with TESE in germinal failure. 42 non-obstructive azoospermic patients revealed no spermatozoa at all in their ejaculates, even after centrifuge. The histology of 42 patients revealed 15 Sertoli cell only Syndrome, 4 maturation arrest and 23 severe hypospermatogenesis. All patients underwent extensive multiple testicular biopsy for sperm retrieval. These patients were scheduled for ICSI using testicular spermatozoa. In 25 out of 42 non-obstructive azoospermic patients, spermatozoa were recovered from multiple testicular biopsy specimen and 11 ongoing pregnancies were achieved. There are usually some tiny foci of spermatogenesis which allow TESE with ICSI in non-obstructive azoospermia. Also these patients may have sufficient sperm in the testes for ICSI, despite extremely high FSH level and small testes.
Azoospermia* ; Biopsy ; Humans ; Male ; Oligospermia ; Pregnancy ; Sertoli Cell-Only Syndrome ; Sperm Injections, Intracytoplasmic ; Sperm Retrieval ; Spermatogenesis ; Spermatozoa ; Testis

PURPOSE: This study investigated the relationship between spermatogenesis and telomerase activity in sperm pellet and testicular biopsy specimen to define the pathophysiology. MATERIALS AND METHODS: PCR-based telomeric repeat amplification protocol(TRAP) assay was used to detect telomerase activity of sperm pellet from 10 normal fertile volunteers, 25 sperm pellet from infertile patients with oligospermia. and 48 testicular tissues from biopsy or orchiectomy followed by various causes. The telomerase activity were compared with the Johnsen score and serum hormonal levels. RESULTS: Any Telomerase activity was not detected in sperm pellet from normal volunteers and 22 of 25 (88%) sperm pellet from infertile patients with oligospermia. In testicular tissues, telomerase activity was expressed in all patients with normal spermatogenesis, while was not expressed in all with Sertoli cell only syndrome. Of one disorganization & sloughing, 15 hypospermatogenesis and 6 maturation arrest, one (100%), 14 (93.3%) and 5 (83.3%) cases expressed telomerase activity. Frequency and density of telomerase activity were increased according to the increasing of Johnsen score, but no statistical significance. Telomerase activity was not related with serum hormone levels including FSH, LH, T, E2 and prolactin. CONCLUSIONS: Telomerase appears to be present in testicular tissue that harbors immature germ cells and is absent in tissue that contains only Sertoli cells. These data indicate that measurement of telomerase activity may be a useful predictor of spermatogenic activity for applicating the assisted reproductive technology including testicular sperm extraction in male infertile patient with Sertoli cell only syndrome.
Biopsy ; Germ Cells ; Healthy Volunteers ; Humans ; Male ; Oligospermia ; Orchiectomy ; Prolactin ; Reproductive Techniques, Assisted ; Sertoli Cell-Only Syndrome ; Sertoli Cells ; Spermatogenesis* ; Spermatozoa ; Telomerase* ; Volunteers

Electron microscopic observation of fine structural changes of Sertoli cells in the male sterility patients was made on 10 histologically proved cases. They were histologically classified to 3 groups by Nelson's classification, hypospermatogenesis, spermatogenic arrest and germinal cell aplasia. The results obtained were summarized as follows. l) The findings of ultrastructures of Sertoli cells in the normal man showed no significant differences with other author's results. 2) The ultrastructures of Sertoli cells in the male sterility revealed degenerative changes, characterized by fragmentation and dilatation of subsurface cisternae, swelling and vacuolization of mitochondria with decrease in the number of cristae and poor development of Golgi apparatus. And also observed were proliferative changes, characterized by increase in the number of sER, rER and ribosome. These morphological changes were severer in spermatogenic arrest and the severest in germinal cell aplasia. 3) The results were suggested that the most significant ultrastructural changes of Sertoli cells of male sterility were the degenerative changes of the organelles of protein synthesis and protein transport system such as rER, ribosome and Golgi apparatus.
Classification ; Dilatation ; Golgi Apparatus ; Humans* ; Infertility ; Infertility, Male* ; Male ; Male* ; Mitochondria ; Oligospermia ; Organelles ; Protein Transport ; Ribosomes ; Sertoli Cell-Only Syndrome ; Sertoli Cells*

OBJECTIVETo establish a stable and reliable model of Sertoli-cell-only syndrome in mice.

METHODSWe randomly divided 60 NIH mice into two groups of equal number to receive intraperitoneal injection of busulfan (30 mg/kg) and 30 or 60 minutes of testis cooling. At 2, 4 and 8 weeks after treatment, we recorded the survival rate of the mice, weight of the testis and Johnsen scores, and conducted quantitative analysis on the degrees of spermatogenetic failure.

RESULTSThere were no significant differences in the baseline body weight and survival rate between the intervention and control groups (P > 0.05). At 4 and 8 weeks, the testis weight and Johnsen score were significantly lower in the intervention group than in the control ([0.04 +/- 0.01] g and [0.05 +/- 0.01] g vs [0.09 +/- 0.03] g and [0.11 +/- 0.02] g, P < 0.05; 3.86 +/- 0.50 and 2.70 +/- 0.67 vs 9.60 +/- 0.25 and 9.76 +/- 0.43, P < 0.01). At 2, 4 and 8 weeks, the testis weights were (0.07 +/- 0.02) g, (0.06 +/- 0.01) g and (0.09 +/- 0.01) g, respectively, in the 30-min cooling group and (0.05 +/- 0.01) g, (0.04 +/- 0.02) g and (0.04 +/- 0.02) g in the 60-min cooling group, significantly lower than in the control side at the same time points ([0.11 +/- 0.01] g, [0.11 +/- 0.01] g and [0.12 +/- 0.00] g) (P < 0.05), and the Johnsen scores were 4.70 +/- 0.67, 2.70 +/- 0.84 and 6.10 +/- 1.14 in the 30-min and 1.67 +/- 0.58, 1.20 +/- 0.45 and 1.00 +/- 0.00 in the 60-min cooling group, remarkably lower than in the control side (9.60 +/- 3.23, 9.60 +/- 0.55 and 9.70 +/- 0.45) (P < 0.01). Histopathological examination of the cooled testes revealed considerable atrophy of seminal tubules, necrosis of seminiferous epithelia and peritubular fibrosis.

CONCLUSIONAdministration of busulfan has no obvious influence on the survival of mice, and is a reliable method for constructing a mouse model of Sertoli-cell-only syndrome.

Animals ; Busulfan ; adverse effects ; Cold Temperature ; Disease Models, Animal ; Male ; Mice ; Mice, Inbred Strains ; Organ Size ; Sertoli Cell-Only Syndrome ; chemically induced ; Sertoli Cells ; Testis