PURPOSE: The pathogenesis of infertility in unilateral cryptorchidism remains unclear. We studied prospectively to evaluate the cause concerning potential infertility in unilateral inguinal cryptorchidism.Materials and Methods: Between Feb 1998 and July 2000, 30 specimens were taken by ipsilateral undescended and contralateral descened testicular biopsies in 15 unilateral inguinal cryptorchid boys (age range: 1-11 years, mean: 4.7 years). Control testicular biopsies were performed in 5 hydrocele boys (age range: 1-9 years, mean: 5.1 years). We performed histomorphologic analysis including spermatogonia per tubule (S/T) value, Sertoli cell index (SCI), tubular degeneration phase V-VII (TDP V-VII), mean tubular diameter (MTD), and changes of peritubular interstitial tissue (thickened tubular basement membrane and peritubular fibrosis). RESULTS: Testis volume, S/T value, and MTD were significantly different between ipsilateral cryptorchid and contralateral testes. However, there was no significant difference between ipsilateral cryptorchid and contralateral testis in SCI, TDP V-VII, and changes of peritubular interstitial tissue. We found significant difference between contralateral and control testis in testis volume, S/T value, MTD, TDP V-VII, and changes of peritubular interstitial tissue except SCI.Conclusions: Decreased testis volume, S/T value, MTD and increased TDP V-VII of contralateral testis are associated with germinal hypoplasia. These findings may explain the pathogenesis of infertility in unilateral inguinal cryptorchidism.
Basement Membrane ; Biopsy ; Cryptorchidism* ; Infertility ; Male ; Prospective Studies ; Spermatogonia ; Testis*

PURPOSE: Although retractile testes are frequently found in the pediatric population, there are controversies in the management of retractile testes. We investigated the necessity of treatment for retractile testes by analyzing their histologic findings. MATERIALS AND METHODS: Sixty-one testicular biopsies were performed during orchiopexy from 36 boys(range: 1.3-12.9 years, mean: 5.4 years) with retractile testes(11 unilateral, 50 bilateral) and 115 testicular biopsies from 83 cryptorchid patients(range: 0.6-15.0 years, mean: 3.7 years, 51 unilateral, 64 bilateral). Parameters for both Sertoli cell and germ cell were determined in each group. RESULTS: The average tubular degeneration phase(TDP) V-VII were 0.23+/-0.18 for retractile testes and 0.22+/-0.17 for cryptorchid testes and were not statistically different. Both the average sertoli cell index(SCI) and mean spermatogonia per tubules(S/T) value were statistically different between retractile and cryptrochid testes with values of 26.81+/-6.75, 23.04+/-5.85(p<0.01) and 2.96+/-1.33, 0.61+/-0.87(p<0.01), respectively. CONCLUSIONS: Although S/T value of retractile testes was higher than that of cryptorchid testes, Sertoli cell degenerative patterns were similar. These findings might indicate that retractile testis needs treatment like cryptorchid testis does. However, further investigation is warranted to elucidate whether these changes are normal variations since changes are observed in both Sertoli & germ cells in normal boys as they are aging.
Aging ; Biopsy ; Germ Cells ; Orchiopexy ; Pathology ; Spermatogonia ; Testis*

The effects of both hyperthermia alone and X-ray irradiation combined with hyperthermia on rat testis have been investigated. The histological changes were observed on 15 and 30 days after treatment. There was no histological change of rat testis by hyperthermia alone. The earliest change by x-ray irradiation was the degeneration of the spermatogonia of the seminiferous tubule, which was appeared in 2 gy group. Necrosis of the spermatogonia was severe in 6 gy group and complete atrophy was developed in 8 gy group. With increased dose of radiation, the degrees of changes of tubules was increased. In combined group of X-ray irradiation and hyperthermia, the histological change of the seminiferous tubule was more severe than X-ray alone group. Necrosis and atrophy of the spermatogonia were appeared in 2 gy and complete atrophy of spermatogonia was seen in 6 gy group. Thermal enhancement ratio (calculated at the complete atrophy of the spermatogonia) was 1.3 in this experiment. There was no difference in observation time inverval between 15 and 30 days after each treatment in all groups.
Animals ; Atrophy ; Fever* ; Necrosis ; Rats* ; Seminiferous Tubules ; Spermatogonia ; Testis*

Adult Germline Stem Cells ; Humans ; Male ; Mutation ; Spermatogenesis ; Spermatogonia

OBJECTIVETo establish a long-term culture system for mouse spermatogonial stem cells (SSCs) and to discuss the key factor that supports mouse SSC self-renewal and proliferation.

METHODSTestis cells from 4-6 days postpartum male transgenic BALB/c mce were collected by a modified two-step enzymatic digestion method and plated on 0. 2% elatin-coated tissue culture plates. The germ cells were enriched by differential adherence selections after respectively incubated for 1, 5 and 24 h and then plated on the mitomycin C-inactivated mouse embryonic fibroblast (MEF) feeder layer. The basal culture medium was StemPro-34 SFM supplemented with other 15 nutrient factors. The 20 ng/ml Glial cell line-derived neurotrophic factor (GDNF), 10 ng/ml basic fibroblast growth factor (bFGF) and 200 ng/ml GDNF-family receptor alpha 1 (GFRalpha1) were added to the serum-free medium to promote SSC proliferation. Several important surface markers and special genes were examined by immunocytochemical staining and RT-PCR analysis.

RESULTSAfter 3-4 days culture on the MEF feeder, SSCs proliferated continuously and formed typical colonies. SSCs from the BALB/c mice could be cultured in a steady state for 3 months. Immunocytochemical staining showed that Oct4 was specifically expressed in the cultured SSC nucleus and GFRalpha1 strongly expressed on the surface of the membrane. RT-PCR confirmed that the cultured SSCs expressed Oct-4, GFRalpha1, Sox2 and several other special genes resembling undifferentiated spermatogonia.

CONCLUSIONSSCs from BALB/c mice could be cultured in the improved culture system for 3 months. This culture system could help further understand the regulating mechanism of SSCs and might provide an opportunity for the treatment of male infertility by SSC transplantation.

Animals ; Cell Culture Techniques ; methods ; Male ; Mice ; Mice, Inbred BALB C ; Spermatogonia ; cytology ; Stem Cells ; cytology

The isolation and culturation of SSCs of different stage of Wuzhishan Mini Porcine (WZSP) with different way of enzymatic digestion and culturation were deaded in this study. The results of the experiment described are as the following: The proper time of isolation and culturation of SSCs of WZSP is 1-20 old days. Different old of piglets with different method. Using DMEM medium as a fundmental culture medium add different gradient at 34 degrees C in a water-saturated atmosphere of 95% air, 5% CO2. The mulberry-shaped SSCs clusters appeared as original generation in 7-8 days culture. The SSCs clusters developed half-suspendedly in the culture medium. SSCs alkaline phosphatase (AKP) staining expressed positively. Mouse embryonic fibroblast was used as feeder layer for the SSCs passage cultured, The SSCs show good attached attributes, but the number of SSCs decreased quickly after 4 days culture. By seminiferous cord fragment culturation can also appear SSCs clusters in 5 days, The SSCs clusters developed half-suspendedly in the culture medium. In addition, the testes placed in cold (4 degrees C) PBS banlanced salt solution for 24 h also can be used as a good matierials for preparation of SSCs. These results indicate that the method of solation and culturation of SSCs are very correct and efficient, all these can be utilized as a good reference for future studies.
Animals ; Cells, Cultured ; Male ; Mice ; Spermatogonia ; cytology ; Stem Cells ; physiology ; Swine

The self-renewal and differentiation of adult stem cells are closely related to their niches. Naturally, spermatogonial stem cells (SSCs) are the only adult stem cells in the body, which can transfer genetic information into the offspring. An insight into the modulation of the self-renewal and differentiation of SSCs can help elucidate the mechanisms of spermatogenesis and investigate the proliferation and differentiation of other adult stem cells. Therefore, the SSC system provides an ideal model for researches on the adult stem cell niche. More and more evidence indicates that the self-renewal and differentiation of SSCs are regulated by their niches. Based on our previous work and other related findings recently reported, this article presents an overview on the biological properties of SSC niches and their relationship with the self-renewal and differentiation of SSCs, focusing on the basic properties and components of SSC niches and various regulatory factors they produce.
Animals ; Cell Differentiation ; Male ; Spermatogenesis ; Spermatogonia ; cytology ; Stem Cell Niche ; Stem Cells ; cytology

OBJECTIVESTo detect the changes of DNA ploidy of spermatogenic cells in testis and epididymis.

METHODSRight epididymides and testes from 15 fertile youth donors who died of accident were collected. Samples of spermatogenic cells in different regions of epididymis (caput, corpus and cauda) and tests were collected. DNA of spermatogenic cells were detected by flow cyctometry (FCM).

RESULTSThe haploid(1n), diploid(2n) and tetraploid(4n) spermatogenic cells were existed in different regions of epididymis and testis. The 1n cells increased from (24.87 +/- 7.28)% in testis to (96.33 +/- 1.58)% in epididymis cauda, there were significant differences among regions of testis and epididymis caput, corpus(P < 0.01), and the difference among regions of epididymis corpus and epididymis cauda were also significant(P < 0.05). While the percentages of 2n and 4n cells decreased from (63.07 +/- 8.96)% and (9.43 +/- 3.83)% in tesits to (2.47 +/- 0.93)% and (1.17 +/- 0.95)% in epididymis respectively. There was significant difference of 2n cells between testis and epididymis caput, corpus(P < 0.01), and was also remarkable difference between epididymis corpus and cauda (P < 0.05). There was no difference of 4n cells between testis and epididymis caput(P > 0.05). There were significant difference among regions of epididymis caput, corpus and cauda(P < 0.05).

CONCLUSIONSThe percentage of immature spermatogenic cells decreased along with passing through the epididymis.

Adult ; DNA ; analysis ; Epididymis ; metabolism ; Flow Cytometry ; Humans ; Male ; Spermatogonia ; metabolism ; Testis ; metabolism

OBJECTIVETo study the expression of Annexin A7 in the mouse testis, especially in different types of spermatogonia.

METHODSWe prepared Annexin A7 recombinant protein using prokaryotic expression, adsorbed the Annexin A7 antibody with it after identified by mass spectrometry, and detected the expression of Annexin A7 by Western-blot and immunohistochemistry.

RESULTSAnnexin A7 was expressed in a development-dependent manner in the spermatogonia of the prepubertal mice and in the type-A single (As) and type-A paired (Apr) spermatogonia of adult mice. These results were confirmed by the co-localization of Annexin A7 and Stra8, a known determinant of differentiated spermatogonial stem cells (SSCs).

CONCLUSIONAnnexin A7 is the internal factor of As and Apr spermatogonia, which might be involved in the biological functions of SSCs.

Animals ; Annexin A7 ; metabolism ; Male ; Mice ; Spermatogonia ; cytology ; metabolism ; Stem Cells ; cytology ; metabolism

Undescended testis is one of the most common anomalies of genitourinary tract in children but optimal time for treatment of it has not been determined still. We examined the changes of seminiferous tubules according to ages in 41 patients with undescended testis which orchiectomy or testicular biopsy were performed in the Department of Urology, Kyung Hee University Hospital during the period from January, 1979 to April, 1985 and following results were obtained. l. The ages of 41 patients ranged from 6 years to 48 years. Of these,36 patients were unilateral and 5 patients were bilateral undescended testis. 41 undescended testes were located in inguinal canal, 2 in high scrotum and 3 above the internal inguinal ring. 2. Mean Tubular Diameter was average 48.5 Um from 6 years to 12 years, 81.2 Um at 14 years and 118.4 Um at 24 years. But in normal group, it was average 95.7 Um from 6 years to 12 years, 2l3.6 Um at 14 years and 229.4 Um at 17 years. So, there was already severe damage in development of seminiferous tubules in undescended testis at 6 years. 3. Mean Tubular Fertility Index was average 19.7% from 6 years to 17 years, 42% at 19 years, average 10.8% from 20years to 30 years and 0% at 48 years. But in normal group, it was 78.0% at 6 years and 100% at I4 years. So. there was already severe damage in formation of spermatogonia in undescended testis at 6 years. 4. The thickness of basement membrane of seminiferous tubules in undescended testis was 2.2 Um at 6 years and increased with age to 9.9 Um at 24 years.
Basement Membrane ; Biopsy ; Child ; Cryptorchidism* ; Fertility ; Humans ; Inguinal Canal ; Male ; Orchiectomy ; Scrotum ; Seminiferous Tubules ; Spermatogonia ; Urology