Faculty Opinions recommendation of Vitrification with microinjection of single seminiferous tubules: an efficient cryopreservation approach for limited testicular tissue.

2022 ◽  
Author(s):  
Ralf Henkel
Keyword(s):  
Testicular Tissue ◽  
Seminiferous Tubules

scholarly journals The Expression Levels and Cellular Localization of Pigment Epithelium Derived Factor (PEDF) in Mouse Testis: Its Possible Involvement in the Differentiation of Spermatogonial Cells

2021 ◽  
Vol 22 (3) ◽  
pp. 1147
Author(s):  
Noy Bagdadi ◽  
Alaa Sawaied ◽  
Ali AbuMadighem ◽  
Eitan Lunenfeld ◽  
Mahmoud Huleihel
Keyword(s):  
Cellular Localization ◽  
Pigment Epithelium ◽  
Testicular Tissue ◽  
Seminiferous Tubules ◽  
Target Cells ◽  
Isolated Cells ◽  
Expression Levels ◽  
Cellular Origin ◽  
Pigment Epithelium Derived Factor

Pigment epithelium derived factor (PEDF) is a multifunctional secretory soluble glycoprotein that belongs to the serine protease inhibitor (serpin) family. It was reported to have neurotrophic, anti-angiogenic and anti-tumorigenic activity. Recently, PEDF was found in testicular peritubular cells and it was assumed to be involved in the avascular nature of seminiferous tubules. The aim of this study was to determine the cellular origin, expression levels and target cells of PEDF in testicular tissue of immature and adult mice under physiological conditions, and to explore its possible role in the process of spermatogenesis in vitro. Using immunofluorescence staining, we showed that PEDF was localized in spermatogenic cells at different stages of development as well as in the somatic cells of the testis. Its protein levels in testicular homogenates and Sertoli cells supernatant showed a significant decrease with age. PEDF receptor (PEDF-R) was localized within the seminiferous tubule cells and in the interstitial cells compartment. Its RNA expression levels showed an increase with age until 8 weeks followed by a decrease. RNA levels of PEDF-R showed the opposite trend of the protein. Addition of PEDF to cultures of isolated cells from the seminiferous tubules did not changed their proliferation rate, however, a significant increase was observed in number of meiotic/post meiotic cells at 1000 ng/mL of PEDF; indicating an in vitro differentiation effect. This study may suggest a role for PEDF in the process of spermatogenesis.

Consequences of Microwave oven Exposed Diet on The Basal Lamina of Seminiferous Tubules of Mice

2021 ◽  
Vol 15 (8) ◽  
pp. 2141-2144
Author(s):  
Kishwar Naheed ◽  
Muhammad Saad Abdullah ◽  
Maria Yousaf ◽  
Humaira Ali ◽  
Fareeha Mushtaq ◽  
...  
Keyword(s):  
Basal Lamina ◽  
Treated Group ◽  
Testicular Tissue ◽  
Microwave Oven ◽  
Mentha Piperita ◽  
Seminiferous Tubules ◽  
Control Group ◽  
Protective Effects ◽  
Trial Methodology ◽  
Experimental Group

Usage of electronic gadgets like microwave oven is increasing day by day that heats the food by exposing it to electromagnetic radiations which has many hazardous effects on human health including fertility. Aim: To find the effects of microwave oven exposed diet on basal lamina of seminiferous tubules of mice alongwith protective effects of Mentha piperita and melatonin on the same tissue. Study Design: Randomized control trial. Methodology: Adult male mice (n=32) were divided into four groups. Control group (G1) received standard pellets prepared for mice. Second group (G2) was given mice pellets exposed to microwave oven. Third group (G3) received Mentha Piperita leaf extract along with mice pellets exposed to microwave oven and the fourth group (G4) received oral melatonin along with pellets exposed to microwave oven. Later their testicular tissue was removed for histological examination while basal lamina disruption was assessed by scoring. Data analyzed by SPSS 22.0v. Results: In group G2, there was slight disruption in the basal lamina in 75% of the cases while in experimental group G3, there was slight disruption of basal lamina only in 12.5% of the cases. However, in group G4, only 25% specimen had slight disruption of basal lamina Conclusion: It was concluded that microwave oven exposed diet produced severe disruption of basal lamina in group G2 that decreased in Mentha piperita and melatonin treated groups. However, Mentha piperita treated group produced better results than melatonin treated group. Keywords: Mice, Testis, Basal Lamina, Mentha piperita and Melatonin

scholarly journals The Effects of Vasectomy on Testicular Tissue of Mice: Histological Changes and DNA Fragmentation Study

2020 ◽  
Vol 11 (2) ◽  
Author(s):  
Salman MO ◽  
Al-Wasiti EA ◽  
Thamir KA ◽  
Al-Ani IM ◽  
Al-Salihi AR
Keyword(s):  
Single Cell ◽  
Dna Fragmentation ◽  
Gel Electrophoresis ◽  
Testicular Tissue ◽  
Seminiferous Tubules ◽  
Control Group ◽  
Sham Operation ◽  
Male Mice ◽  
Histological Changes ◽  
Single Cell Gel Electrophoresis

Introduction: We aim to investigate the effect of vasectomy on the histology of the testis as well as to evaluate DNA fragmentation in testicular tissue of male mice. Methods: Bilateral vasectomy was performed on 20 mature male mice; 10 control mice underwent sham-operation. After 6 weeks, the testes were evaluated for histological changes and DNA fragmentation by single cell gel electrophoresis (comet assay). Results: Marked alterations were observed in the testes of vasectomized mice, including degeneration of spermatids, thickened basement membrane, dilatation of the seminiferous tubules, exfoliation of germ cells, reduction in the seminiferous cell population, vacuolated appearance of the epithelium in the tubules and marked interstitial fibrosis. Single cell gel electrophoresis showed a highly significant (P<0.0001) increase in DNA damage among vasectomized mice (46.02%) compared with control group (%27.17) after six weeks of operation. Conclusion: Vasectomy induced deterioration in the seminiferous tubules associated with increased testicular cell’s DNA fragmentation.

P–056 To graft or not to graft? Intratesticular grafting of testicular tissue from Klinefelter boys to the mouse testis as possible novel in vivo model

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
M Willems ◽  
P Sesenhausen ◽  
I Gies ◽  
V Vloeberghs ◽  
J D Schepper ◽  
...  
Keyword(s):  
Klinefelter Syndrome ◽  
Testicular Biopsy ◽  
Testicular Tissue ◽  
Mouse Testis ◽  
In Vivo Model ◽  
Seminiferous Tubules ◽  
Positive Effects ◽  
Fibrotic Process

Abstract Study question Can intratesticular transplanted testis tissue from Klinefelter boys to the mouse testis be used to study the mechanisms behind testicular fibrosis? Summary answer Grafting of testicular tissue from Klinefelter boys to the mouse testis is not a valuable new in vivo model to study Klinefelter-related testicular fibrosis. What is known already Klinefelter syndrome (KS; 47, XXY) affects 1–2 in 1000 males. Most KS men suffer from azoospermia due to a loss of spermatogonial stem cells. Additionally, testicular fibrosis is detected from puberty onwards. However, mechanisms responsible for fibrosis and germ cell loss remain unknown. An optimal in vivo model to study the KS testicular fibrotic process is not available. This study aimed to evaluate a possible in vivo model to study KS-related testicular fibrosis. In addition, the effect of the mast cell blocker ketotifen, which showed positive effects on fertility in infertile non-KS patients, was evaluated in this graft model. Study design, size, duration First, the survival time of the KS graft was established, since it was the first time KS tissue was transplanted to the mouse testis. Testes were collected after two, four, six and eight weeks after which histological and immunohistochemical evaluations were performed. Next, the effect of daily ketotifen injections on the fibrotic appearance of intratesticular grafted testicular tissue from KS and controls was evaluated. Participants/materials, setting, methods Testicular biopsy samples from pre- and peripubertal KS (n = 22) and age-matched control samples (n = 22) were transplanted to the testes of six weeks old Swiss Nu/Nu mice (n = 22). Prior to grafting, testicular tissue pieces were cultured in vascular endothelial growth factor (VEGF) for five days. Next, tissues were transplanted to the mouse testes. Testicular transplants were analysed by immunohistochemistry. In the second experiment, mice were given daily subcutaneous injections of ketotifen or saline. Main results and the role of chance Four weeks after transplantation, all KS grafts could still be retrieved. At a later timepoint, degeneration of the tissue could be detected. In the grafts, recovered four weeks after transplantation, about 30% of the tubules in peripubertal grafts showed a good integrity, while in the prepubertal tissue, 83% of the tubules were intact. A fibrotic score was assigned to each graft. No significant changes in fibrotic score was observed between testicular biopsies before or after transplantation. However, an increased (p &lt; 0.01) fibrotic score was observed after in-vitro treatment with VEGF both in control and KS tissue. Based on recovery and tubule integrity grafts were recovered after four weeks in the second experiment. Treatment with ketotifen did not result in significant histological differences compared to non-treated grafts (KS and control tissue). The survival potential of grafts from KS testicular biopsies of pre- and peripubertal boys was patient- and age-dependent. After four weeks, most KS tissue starts to degenerate. In prepubertal tissue, seminiferous tubules were mostly intact, while tissue from adolescent boys was impaired. Interestingly, no loss of germ cells was observed after transplantation of the testicular tissue. Limitations, reasons for caution The availability of tissue from young KS patients is very scarce, leading to a low number of included patients (n = 8). Testicular tissue pieces from the same patient were included to evaluate the differences before and after transplantation. However, histological variability between testicular tissue biopsy pieces is well-known in KS patients. Wider implications of the findings Since testicular tissue from KS boys, transplanted to the mouse testes, already starts to degenerate after four weeks and the integrity is not optimal, we conclude that this is not a valuable model for future studies. In vitro models to study the KS-testicular fibrosis should be investigated. Trial registration number NA

scholarly journals Effects of microgravity or simulated launch on testicular function in rats

1992 ◽  
Vol 73 (2) ◽  
pp. S174-S185 ◽  
Author(s):  
R. P. Amann ◽  
D. R. Deaver ◽  
B. R. Zirkin ◽  
G. S. Grills ◽  
W. J. Sapp ◽  
...  
Keyword(s):  
Germ Cells ◽  
Quantitative Methods ◽  
Smooth Endoplasmic Reticulum ◽  
Testicular Tissue ◽  
Seminiferous Tubules ◽  
Specific Gene ◽  
Fixed Tissue ◽  
Treatment Groups ◽  
Essentially Normal ◽  
Peripheral Blood Plasma

Testes from flight rats on COSMOS 2044 and simulated-launch, vivarium, or caudal-elevation control rats (5/group) were analyzed by subjective and quantitative methods. On the basis of observations of fixed tissue, it was evident that some rats had testicular abnormalities unassociated with treatment and probably existing when they were assigned randomly to the four treatment groups. Considering rats without preexisting abnormalities, diameter of seminiferous tubules and numbers of germ cells per tubule cross section were lower (P less than 0.05) in flight than in simulated-launch or vivarium rats. However, ratios of germ cells to each other or to Sertoli cells and number of homogenization-resistant spermatids did not differ from values for simulated-launch or vivarium controls. Expression of testis-specific gene products was not greatly altered by flight. Furthermore, there was no evidence for production of stress-inducible transcripts of the hsp70 or hsp90 genes. Concentration of receptors for rat luteinizing hormone in testicular tissue and surface density of smooth endoplasmic reticulum in Leydig cells were similar in flight and simulated-launch rats. However, concentrations of testosterone in testicular tissue or peripheral blood plasma were reduced (P less than 0.05) in flight rats to less than 20% of values for simulated-launch or vivarium controls. Thus spermatogenesis was essentially normal in flight rats, but production of testosterone was severely depressed. Exposure to microgravity for greater than 2 wk might result in additional changes. Sequelae of reduced androgen production associated with microgravity on turnover of muscle and bone should be considered.

25 Cryopreservation of horse testicular tissue as a model for rhinoceros

2019 ◽  
Vol 31 (1) ◽  
pp. 138 ◽  
Author(s):  
M. C. Gómez ◽  
A. Alrashed ◽  
C.-Y. Su ◽  
B. Durrant
Keyword(s):  
Basement Membrane ◽  
Dimethyl Sulfoxide ◽  
Structural Integrity ◽  
Membrane Damage ◽  
Genetic Material ◽  
Testicular Tissue ◽  
Live Cells ◽  
Seminiferous Tubules ◽  
Positive Interaction ◽  
Cryoprotectant Solution

Cryopreservation of testicular tissue (TT) allows retention of valuable genetic material that can be used for conservation of endangered species, such as the northern white rhinoceros (NWR; Ceratotherium simum cottoni). Previously, we found that cryopreservation of NWR TT with a slow controlled cooling rate (CR) method induced morphological alterations in the seminiferous tubules (ST). However, the relative influence of CR, type of medium, and condition of TT from the aged NWR male on TT integrity was not clear. Due to the limited availability of rhinoceros TT, we used the horse as a model for optimization of TT cryopreservation. We evaluated the effect of (1) cryoprotectant solution [PBS (PBS +1.5M dimethyl sulfoxide) v. DMEM (DMEM/F12+10.0% fetal bovine serum+0.05M sucrose+1.5M dimethyl sulfoxide)] and (2) CR [CR1 (−2.0°C min−1 from 0°C to −4.0°C, −15°C min−1 to −12°C, and −0.3°C min−1 to −40°C in a programmable freezer) v. CR2 (same as CR1 but cooled to −8°C and held for 5min before cooling to −40°C) v. CR3 (−1.0°C min−1 from 0°C to −80°C in a CoolCell® freezing device; Corning, Corning, NY, USA)] on the structural integrity of ST from a 2-year-old horse (n=20 ST), cell viability, and expression of spermatogonial stem cells (SSC; GFRα1, and GRP125) and pluripotent markers (SSEA-4, SSEA-1, and OCT-4) in spermatogonial cells isolated from TT frozen with the above treatments (n=3). We found a positive interaction between CR and cryoprotectant solution on structural integrity of fixed and stained TT after freezing in PBS and CR2 that resulted in lower detachment of epithelium cells from the basement membrane (score±standard error of the mean; 0.50±0.1) than that of TT frozen in PBS and CR1 and CR3 (1.00±0.1 and 1.80±0.1, respectively; P&lt;0.001) or in DMEM and CR1 (1.25±0.1), CR2 (1.35±0.1), and CR3 (1.40±0.1; P&lt;0.01) and in lower incidence of basement membrane damage (0.75±0.1) than that of TT frozen in PBS and CR1 (1.17±0.07) and CR3 (1.16±0.07) or in DMEM and CR1 (1.10±0.1), CR2 (1.15±0.1), or CR3 (1.45±0.1; P&lt;0.01). A lower rate of pyknosis was observed in TT frozen with PBS (1.15±0.06) than in TT frozen in DMEM (1.43±0.06; P&lt;0.001). Overall, integrity of ST was improved when TT was frozen in PBS at CR2 having similar percentages of ST with intact epithelium (60%) and basement membrane (35%) as that of refrigerated TT (45 and 50%, respectively) but different from that of TT frozen with PBS at CR1 (10 and 15%, respectively; P&lt;0.05). Flow cytometry analysis of spermatogonial cells revealed that the percentages of live cells from TT frozen in PBS (CR1: 61.5±7.4%; CR2: 59.7±4.8%; CR3: 51.5±4.1%) or DMEM (CR1: 66.2±6.0%; CR2: 59.8±6.0%; CR3: 58.9±6.9%), and expression of SSC and pluripotent markers was similar among all freezing treatments. However, the percentages of live cells from frozen-thawed TT were lower than those of cells isolated from refrigerated TT (80.6±2.2%; P&lt;0.001). Overall, our results showed that (1) structural integrity of horse ST was better maintained when TT was frozen in PBS at CR2 and (2) SSC can be isolated from frozen-thawed TT with a similar relative frequency to that of refrigerated TT.

scholarly journals Formalin Versus Bouin Solution for Testis Biopsies: Which Is the Better Fixative?

2020 ◽  
Vol 13 ◽  
pp. 2632010X1989726 ◽  
Author(s):  
James L Ellenburg ◽  
Peter Kolettis ◽  
Joseph C Drwiega ◽  
Anna M Posey ◽  
Matthew Goldberg ◽  
...  
Keyword(s):  
Basement Membrane ◽  
Nuclear Membrane ◽  
Cytoplasmic Membrane ◽  
Testicular Tissue ◽  
Seminiferous Tubules ◽  
High Quality ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
Histologic Evaluation

Purpose: Some resources recommended Bouin solution for the fixation of testis biopsy specimens. We compared the histologic quality of rat testicular tissue using buffered formalin and Bouin solution as fixatives. Methods: We prospectively compared the histologic quality of rat testicular tissue fixed in Bouin solution versus formalin. Testicular tissue was harvested post-mortem from six rats. Each testis was removed and sectioned in half; one half was fixed in formalin and one half in Bouin solution. Testicular tissue histology (nuclear membrane detail, nuclear granularity, cytoplasmic granularity, cytoplasmic membrane detail, and basement membrane detail) was graded as high quality (2) or low quality (1). Sloughing of cells into the lumens of the seminiferous tubules was graded on a 0-3 scale (0=none, 1=mild, 2=moderate, 3=extensive). Results: All slides regardless of fixative were of appropriate quality for the histologic evaluation of spermatogenesis. The average sloughing score for formalin cases was 1.4 and for Bouin cases 1.6. Formalin fixed tissue was found to have high quality nuclear membrane detail (2), nuclear granularity (1.9), and basement membrane detail (2). Cytoplasmic granularity was of lesser but adequate quality (1.4). Cytoplasmic membrane detail was poor, (1). Tissue fixed with Bouin solution had high quality basement membrane detail (2) and adequate cytoplasmic granularity (1.5), nuclear membrane detail (1.3) and nuclear granularity (1.4). Cytoplasmic membrane detail was poor (1). Conclusion: Compared to Bouin solution, formalin fixation of rat testicular tissue produced adequate histology for the evaluation of spermatogenesis and may be superior to Bouin solution for certain cytologic features.

The influence of level of feed intake on sperm-producing capacity of testicular tissue in the ram

1978 ◽  
Vol 29 (1) ◽  
pp. 173 ◽  
Author(s):  
CM Oldham ◽  
NR Adams ◽  
PB Gherardi ◽  
DR Lindsay ◽  
JB Mackintosh
Keyword(s):  
Feed Intake ◽  
Testicular Tissue ◽  
Seminiferous Tubules ◽  
Feeding Period ◽  
Sperm Production

The experiment examined the effect of supplementing a roughage diet with lupin grain on the capacity of testicular tissue to produce spermatozoa. Changes in testicle size were estimated by comparative palpation. At the end of the experiment the rams were castrated and the morphology of the testes and their capacity to produce sperm were studied. At the highest level of feed intake, liveweight increased by 32% and testicle volume by 67% during the feeding period of 9 weeks. Rams on a diet that reduced testicle size produced 18 x l06 sperm/g testis per day, and those on a diet that increased testicle size produced 26 x l06 sperm/g testis per day. Those rams whose testes were increasing in size at castration had significantly larger seminiferous tubules, which occupied a significantly greater proportion of the testicle volume, than rams fed on diets that reduced the size of their testes. The variation in rates of sperm production, together with the large differences in testicle weight, resulted in wide differences in total sperm production among rams fed on different diets.

Ultrastructural And Histopathological Investigation Of Seminiferous Tubules Obtained From Azoospermia Cases In The IVF Center

2020 ◽  
pp. 1-3
Author(s):  
◽  
◽  
◽  
Keyword(s):  
Testicular Biopsy ◽  
Testicular Tissue ◽  
Seminiferous Tubules ◽  
Testicular Sperm Extraction ◽  
Interstitial Tissue ◽  
Testicular Sperm ◽  
Electron Microscopic ◽  
Important Indicator ◽  
Serum Hormone ◽  
Histopathological Results

Objective: In this study we purposed to explore seminiferous tubules via histopathological and electron microscopic methods in testicular biopsy samples obtained TESE and the relationship between the findings and levels of serum FSH, LH, testosterone hormones. Methods: Azoospermia patients were divided into two groups, a positive testicular sperm extraction (TESE(+)) and a negative testicular sperm extraction (TESE(-)). Testicular tissue from biopsy samples were subjected to the light and electron microscopic tissue preparations. Serum hormone levels of patients were determined and analyzed statistically between the groups. Results: Compared the groups, more remarkable damages were detected in the seminiferous tubulus of no sperm group in the light and electron microscopic examinations. Although inflammation and partly tubule degeneration was observed, spermatogenesis and sperm cells were determined in the tubules of sperm pozitive group. In the light and ultrastructural analysis of negative sperm group, macrophages and mast cells in the interstitial tissue, vacuolization of seminiferous tubules, lipid inclusions and Sertoli cell only syndrome were the significant findings. When analyzed serum FSH, LH and testosterone hormones between the groups, FSH and LH hormones were statistically significant while Testosteron hormone was not significant. Conclusion: As a result in the seminiferous tubules of individuals histopathological results revealed that FSH and LH are important indicator of testicular function but Testosteron has not any effect. It was concluded that high levels of these hormones impair spermatogenesis and cause testicular failure.

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