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*

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*

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*

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

Spermatogonial transplantation (ST) is a novel technique which mechanism is similar to other transplantations. The donor testicular cells transferred into recipient testes by the microinjection can initiate spermatogenesis and produce sperm in the recipient testis. The homograft has got success. The xenograft in which (man, horse, bull or rat, can be the donor; immunodeficiency mouse can be the recipient) and cryopreserved germ cells can not clonize in the recipient testes. The reason may be immunoreaction. ST technique will be propitious to these fields, 1. to investigate fundamental aspects of spermatogenesis; 2. to regenerate spermatogenesis in infertile individuals; 3. to develop transgenic animals.
Animals ; Animals, Genetically Modified ; Humans ; Male ; Spermatogenesis ; physiology ; Spermatogonia ; transplantation ; Transplantation, Heterologous

OBJECTIVESTo study the expression and the significance of telomerase gene hTERT in testes of infertile male.

METHODSBy using in situ hybridization(ISH) techniques, the expression of telomerase gene hTERT mRNA in testes of 47 infertile male and 10 normal testicular tissues were observed.

RESULTSIn male testes, there was a positive correlation between the expression of hTERT and the quantity and density of germ cells(spermatogonia, spermatocyte, spermatid). The expression of hTERT in some germinal cell of maturation arrest patients were not significantly different with those of normal.

CONCLUSIONSOur results suggest that the deficiency of telomerase might be a factor for germinal cell maturation arrest and there might be some other etiological factors in these patients. Our study provides experimental groundwork for the gene therapy of male infertility.

Humans ; Infertility, Male ; enzymology ; Male ; Spermatids ; Spermatocytes ; Spermatogenesis ; Spermatogonia ; Telomerase ; deficiency ; genetics ; metabolism ; Testis ; enzymology ; physiology

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

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