The aim of the present study was to evaluate gene expression of constitutive and inductive nitric oxide synthase enzymes (cNOS, iNOS) and measurement of nitric oxide metabolite in ovine sertoli cells under heat stress. Testes of ten male lambs were provided from abattoir and then transported to the lab for separating of sertoli cells. Heat stress was challenged in three groups which consist of control (temperature of 32˚C) group, mild heat stress (temperature of 39˚C) and severe heat stress (temperature of 42˚C). The amount of nitrite (nitric oxide metabolite) was significantly higher in 42 ˚C-group than control (p

Ovarian sex cord stromal tumors present with ovarian masses and symptoms resulted from increased hormone secretion. Gynandroblastoma is a rare tumor consisting of two components: granulosa cell tumor and Sertoli or Sertoli-Leydig cell tumor. In this study, a rare case of recurrent gynandroblastoma consisting of Juvenile Granulosa cell tumor and Sertoli cell tumor is reported.

A 29-year-old woman referred to a medical center in Turkmenistan with irregular menstruation and spotting for one year and progressive abdominal pain for the past four months. A mass in the left ovary was reported on CT scan. The patient underwent left salpingo-oophorectomy surgery, and one pathologist diagnosed Granulosa cell tumor, while another diagnosed Androblastoma. 18 months later, the patient underwent laparotomy due to abdominal pain and an intra-peritoneal mass based on MRI. The pathologic diagnosis was Androblastoma. The patient came to Iran to continue treatment and after reviewing the pathology slides of both surgeries, a neoplastic lesion composed of Juvenile Granulosa cell tumor (70%) and well differentiated Sertoli cell tumor (30%) was found, establishing the recurrent Gynandroblastoma.

Inadequate sampling of the tumor specimen and lack of sufficient accuracy in microscopic examination easily lead to the misdiagnosis of Gynandroblastoma. All patients with this tumor should be periodically followed up for several years in terms of recurrence. Furthermore, genetic screening for DICER1 mutation is important.

Successful spermatogenesis requires a series of tightly controlled epigenetic events leads to condensation of sperm chromatin. Through these epigenetic events, expression of transition nuclear proteins (TNPs) and protamines (PRMs) rise to replace with histones. Many epigenetic factors are involved in regulation of these genes. Therefore, evaluation of histone modifications e.g. H3K9ac and H3K9me2, as powerful epigenetic tool in regulation of mentioned genes, can represent better insight into molecular mechanisms of infertility.
The consent was obtained from 60 azoospermic infertile men referred to Royan Institute, Tehran, Iran, according local ethical approval. Then, based on spermogram and pathological features of patients, testes tissue samples were collected from three groups including complete maturation arrest, sertoli cell only syndrome, and hypospermatogenesis (as positive control). Expression of TNPs and PRMs were evaluated using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Besides, chromatin immunopercipitation (ChIP) coupled with real time- polymerase chain reaction was performed to evaluate the incorporation of H3K9ac and H3K9me2 into regulatory regions of mentioned genes.
Findings: There was a significant decrease in expression of TNP and PRM genes in two groups of spermatogenic failure in comparison to positive control. These findings also confirmed by chromatin immunopercipitation data which revealed decreased incorporation of H3K9ac (activating mark), and increased incorporation of H3K9me2 (repression mark) into regulatory regions of mentioned genes in complete maturation arrest and sertoli cell only syndrome groups vs. positive control.
These finding implies significant association of histone modifications with altered expression of sperm chromatin condensing genes and impairment of spermatogenesis in male infertility.

Mammalian spermatogenesis is a complex process of germ cell development within the seminiferous tubules, including the mitotic phase in which type-A spermatogonial stem cells renew themselves. The process of spermatogenesis is regulated not only by gonadotropins, i.e. FSH and LH, but also by the interactions between spermatogenic cells and somatic Sertoli cells (Skinner 1991 and Griswold 1995). Culture systems for recapitulating spermatogenesis in vitro enable us to identify and characterize the factors or genes involved in germ cell proliferation, meiosis and spermatogenesis. In mice, undifferentiated germ cell lines have been established by immortalizing the germ cells with the simian virus 40 large tumor antigens, or with telomerase catalytic component (Fenget al.2002), Type A spermatogonia include a very small number of SSCs and their more numerous differentiating daughter cells. Initial attempts to isolate SSCs started with the isolation of type A spermatogonia and SSC purification. Type A spermatogonia can be obtained in large numbers from young prepubertal bulls, and it is important to note that there are breed differences. Type A spermatogonia isolation can be achieved through mechanical dissociation and enzymatic digestion of the testicular tissue followed by two purification steps, with a final typical bovine type A spermatogonia suspension of 70%. An evaluation for SSC activity using a transplantation assay adapted for bovine SSCs is described. Bovine Type A spermatogonia can be maintained in vitro for short periods (7 to 15 days) with simple culture conditions. However, expansion of SSC can only be achieved under certain conditions such as specially supplemented medium, specific growth factors, and serial sub-culturing for longer periods of time. After expansion, bovine spermatogonia can be cryopreserved while retaining the ability to proliferate and survive (.Aponte&de Rooij 2008). Bovine culture systems require some form of feeder layer to support the germ cells. Autogenous Sertoli cells (Aponte et al. 2006) or embryonic fibroblasts (Oatley etal. 2004) have been used.
Material and Initially, sertoli cells and SSCs were isolated from 3–7-month-old calvesand, and The minced pieces of testis were suspendedin DMEM, which contained 0.5 mgmL−1 collagenase/dispase, 0.5 mgmL−1 trypsin and 0.08 mgmL−1 DNase, for 60 min (with shaking) at 37◦C.). After three washes in DMEM medium and removal of most of the interstitial cells, a second digestion step (45 min at 32◦C) was performed in DMEM by adding fresh enzymes to the seminiferous cord fragments. Separated from the remaining tubule fragments by centrifugation at 30g for 2 min at 37◦C. After filtration through a 70-μm nylon filter, the medium was aspirated, the cells were washed twice and fresh medium was added. Coated plastic dishes were prepared by incubation with a solution of 5μgmL−1 of datura stramonium agglutinin in phosphate-buffered saline (PBS) at 37◦C for 60 min, followed by extensive washing with PBS, supplemented with 0.5% bovine serum albumin The Sertoli cells were isolated using a procedure described by Scarpinoet al. (1998.) with some modifications. The mixed population of the cells obtained byenzymatic digestion was placed on lectin-coated dishes at a concentration of 1.5×105 cells cm−2 and incubated for 1 h at 32◦C in a humidified atmosphere of 5% CO2 in air. After the incubation, the non-adhering cells were collected by being washed twice with medium. After the incubation, the Non-adhering cells were collected by being washed twice with medium. Alternatively, 48 h after being plated on the lectin-coated dishes, the Sertoli cells were detached by ethylenediamine tetraacetic acid (EDTA)–trypsin treatment (0.02% EDTA–0.1% trypsin in Ca2+ and Mg-free PBS) for 5 min at 37◦C, counted and adjusted to desired densities into each well of a 24-well multidish for secondary culture in DMEM at 32◦C in the presence of 10% fetal bovine serum (FBS; Gibco). This method helped isolate the Sertoli cells with more than 95% purity. After the Sertoli cells had been isolated by DSA lectin, the spermatogonia that remained in suspension were collected and kept at 32◦C in a humidified atmosphere and the presence of 10% FBS. More than 95% of the cells were spermatogonial cells. The number of the Sertoli and spermatogonial cells was determined with a hemocytometer. Cell viability was evaluated by means of the dye exclusion test (0.04% trypan blue solution). Two days after the above-mentioned procedure, the Sertoli cells formed a confluent layer, and spermatogonial cells were co-cultured on top of them. Highly purified human FSH and) were added to spermatogonial cell (EGF supplementation: three groups received 4, 12, 20 and 50 ngmL−1of fresh EGF every 3 days for 2 weeks). The diameters and the number of colonies were determined every 3 days after the appearance of the colonies for 2 weeks. An inverted microscope (Zeiss, Germany) was used to determine the number of the colonies, their diameters being measured by ocular grid. The nature of cells in addition to the morphology and the activity of alkaline phosphatase were confirmed through specific markers Oct-4 and vimentin in colonies and Sertoli cells. In the last stage these cells were frozen-melted in the in vitro environment until the purity and strength of biological stem cell is evaluated.
The number and diameter of colonies increased more rapidly in the sertoli coculture, that it agreed with results of Anjamrooz et al., 2006 and Mohamamadi 2010. The numbers of colonies in the EGF-supplemented groups were not significantly different with the control group (P ≤0.05) (table 1-3), that they were Similar to results of Anjamrooz et al.2006 .The diameters of the colonies in the EGF-treated, in4, 12, 20 ngmL−1, had not significantly different with the control group (P ≤0.05) but in 50 ngmL−1 increased the diameters of the colonies slightly (P ≤0.05) that were Similar to results of Anjamrooz et al.2006 and Koruji et al.2009. In the EGF-treated groups, two pathways seem to have affected the spermatogonial cells. In these groups, the number of the colonies was lower than that of the control group.
The results of this study show that spermatogonial stem cells can be separated with high purity from the testicles of calves and also the mutual relationship between spermatogonial stem cells and Sertoli cells and some growth factors can be used to initiate, maintaining the process of spermatogenesis and the enrichment of stem cells during the culture and increasing the purity and viability of them during solidification.

A 25-year-old man referred to our urologist with chief complaint of right testicular pain ofone month duration. Ultrasonography of the testis was performed two times within eight days.The results of ultrasound showed a tumor in the lower pole of the right testicle. In the first ultrasound, tumor size was 7 mm and in the second one it was 7.6 mm. Tumor markers were measured which showed normal alpha-fetoprotein and beta HCG levels. Right orchiectomy was performed.Histological analysis showed that the tumor was consisted of neoplastic cells with eosinophilic cytoplasm and round nuclei, fibrotic stroma and calcific bodies. The incidence of testicular tumors is only 5 cases per 100,000 men; and less than one percent of testicular tumors are related to Sertoli cells. The results of a review study showed that only 61 cases of large cell calcifying Sertoli cell tumor have been reported until 2005. This tumor can produce estrogen which leads to breast enlargement,sexual impotence and acromegaly in men. However, examinations revealed that our patient did not have any of these problems. In most cases this tumor occurs in childhood and patients younger than 20 years of age, but our patient was 25-year-old. LCCSCT is a rare sex cord-stromal tumor with a low malignant potential. A favorable prognosis can be achieved with radical orchiectomy.

Apoptosis or programmed cell death plays an important role in the pathogenesis of many diseases. Diabetes mellitus is a devastating effect on reproductive system of male sexual function in human and animal models and increased apoptosis. Kudzu root is an ISO Flavonin and saponins and often is used as a reducing agent is glucose. Kudzu is able to lower blood glucose in non-insulin dependent and also, by eliminating free radicals that lead to the reduction of oxidative stress. This study was conducted on the effect of Kudzu roots on apoptosis in the testes of rats diabetic.
In this study, 32 male Wistar rats were randomly selected and divided into four groups: control, diabetic rats, diabetic rats treated with Kudzu 100 mg/kg, diabetic rats treated with Kudzu 50 mg / kg. Diabetic by intraperitoneal injection of streptozotocin55 mg/kg was induced. One week after injection, they were treated with a dose Kudzu 50 and 100 mg/kg for five weeks using gavage. Testicular damage by H & E staining and immunohistochemistry and hormonal and Blood biochemical factors were measured.
Findings: Diabetic rats showed a significant increase in apoptosis in the testis. Decrease in seminiferous tubule diameter of diabetic rats, Sertoli cells, sperm count,Insulin and testosterone levels were shown. In rats treated with Kudzu roots it resulted in significant reduction of apoptosis in diabetic rats and significantly increased body weight, plasma Insulin and testosterone levels. It was observed that the oral administration roots of Kudzu increase sperm count, sperm cell of Sertoli cells.
Discussion & The results of this study confirmed the occurrence of apoptosis in diabetes and root Kudzu of the anti-apoptotic effect notes.