gc 1 spermatogonia cells  (Procell Inc)

Journal: eLife

Article Title: Changes in local mineral homeostasis facilitate the formation of benign and malignant testicular microcalcifications

doi: 10.7554/elife.95545

Figure Lengend Snippet: Figure 6. Alkaline phosphatase activity in normal testis and germ cell neoplasia in situ (GCNIS) and calcium and phosphate induced mineralization in vitro. (A) Staining of alkaline phosphatase activity with BCIP/NBT in normal testis (NT) with adjacent tubules containing GCNIS. Scale bars correspond to 1 mm (left picture) and 100 µm (right pictures). (B) Expression levels of the four alkaline phosphatases in normal testis, GCNIS with microlithiasis, and seminoma. ANOVAs with Dunnett’s multiple comparisons test were used as the statistical tests. (C) Mineralization stained with alizarin red in GC1 cells treated with increasing concentrations of calcium and inorganic phosphate (Pi) for 2, 4, or 7 days. (D) Mineralization stained with alizarin red in GC1 cells with calcium and/or phosphate in the presence of the mineralization inhibitor pyrophosphate (PPi) and/or pyrophosphatase (PPA1) that catalyzes the hydrolysis of PPi to Pi for 4 days. (E) Staining of alkaline phosphatase activity with BCIP/NBT in GC1 or TCam2 cells in pH=7 or pH=9.2–9.5. (F) Fast Blue RR staining of alkaline phosphatase activity in GC1 cells cultured in control or osteogenic medium for 14 days.

Article Snippet: GC1 cells (ATCC CRL- 2053, RRID:CVCL_8872) were grown under standard conditions at 37°C at 5% CO2 in DMEM supplemented with penicillin (100 U/ml) and streptomycin (100 mg/ml) (Gibco), 10% FBS (Gibco, #11573397), and L- glutamine (Gibco, #25030–024) (2 mM final conc.) was also added Boisen et al. eLife 2024;13:RP95545.

Techniques: Activity Assay, In Situ, In Vitro, Staining, Expressing, Cell Culture, Control

Journal: Cell Death Discovery

Article Title: Diazinon induces testicular dysfunction and testicular cell damage through increased reactive oxygen species production in mouse

doi: 10.1038/s41420-025-02399-8

Figure Lengend Snippet: A Body weight changes were measured weekly until the end of the experiment (0–11 weeks). Data are presented as the mean ± standard deviation (SD). B Comparison of testicular weight and seminiferous tubule diameter in DZN-injected and control mice. The data is shown as mean ± SD. C H&E staining of testes from DZN-injected and control mice, comparison of positive cells of DDX4 and SYCP3 in mouse testes of each experimental group. Scale bar = 100 μm. ( D ) Number of DDX4+ and SYCP3+ cells in testicular tubules. At least 40 tubules were scored for each testis (five to six biological replicates). Values are presented as the mean ± SD. ( E ) Protein expression levels of SYCP3 and DDX4 in testes of DZN-treated and control mice. Graph represents relative protein levels (mean ± SD, n = 5). ( F ) Expression levels of Sall4, Plzf, and piwii in testes from each group. Values are presented as the mean ± SD (log2 scale). ( G ) Expression levels of Sycp3 , Dmc1 , Rec8 , Pgk2 , and Tp1 . Values on the graph are presented as the mean ± SD (log2 scale). ( H ) Comparison of sperm motility and parameter including distance average path (DAP, μm), distance curved line (DCL, μm), distance straight line (DSL, μm), velocity average path (VAP, μm/s), velocity curved line (VCL, μm/s), velocity straight line (VSL, μm/s) and percentage of fast motility, slow motility, local mortality and immortality through CASA analysis. ( I ) Microscopic analysis of sperm from each experiment group. The graph shows the proportion of abnormal structures in the head, tail, and neck of sperm from DZN-treated and control mice. Abnormality of sperm rations was calculated from seven independent experiments and measured 100 spermatozoa at least in each independent experiment. Asterisks indicate significant differences between the treatment and control groups at *p < 0.05, **p < 0.01, and ***p < 0.001. Scale bar = 50 μm. All statistical analyzes for the graphs were conducted using a t-test.

Article Snippet: Three testicular cell lines, GC-1 spermatogonia (spg), TM3 Leydig, and TM4 Sertoli cells, were purchased from the Korea Cell Bank (KCLB, Seoul, South Korea).

Techniques: Standard Deviation, Comparison, Injection, Control, Staining, Expressing

Journal: Cell Death Discovery

Article Title: Diazinon induces testicular dysfunction and testicular cell damage through increased reactive oxygen species production in mouse

doi: 10.1038/s41420-025-02399-8

Figure Lengend Snippet: A Immunostaining of SOX9, a Sertoli cell marker, in testes of DZN-treated and control mice. Scale bar = 100 µm. B The number of SOX9+ cells in testicular tubules. At least 40 tubules were scored for each testis (five to six biological replicates). Values on graph are presented as the mean ± SD. C Gene expression levels of Sox9 , Amh, and Wt1 in testis from each group. Values on graph are presented as the mean ± SD (log2 scale). D Protein expression levels of SOX9 in testes of DZN-treated and control mice. The graph shows relative protein expression levels (mean ± SD, n = 5). E Immunostaining of HSD3β1 in testes of DZN-treated and control mice. Scale bar = 100 µm. F Concentration of serum testosterone in DZN-treated and control mice. Values on graph are presented as the mean ± SD. G Expression levels of Hsd17b3 , Cyp17a1 , Hsd3b1 , and Cyp11a1 in testis from each group. Values on graph are presented as the mean ± SD (log2 scale) (n = 5). H Protein expression levels of HSD3β1 in testes of DZN-treated and control mice. The graph shows the relative protein levels (mean ± SD, n = 5). Asterisks indicate significant differences between the treatment and control groups at *p < 0.05, **p < 0.01, and ***p < 0.001. All statistical analyzes for the graphs were conducted using a t-test.

Article Snippet: Three testicular cell lines, GC-1 spermatogonia (spg), TM3 Leydig, and TM4 Sertoli cells, were purchased from the Korea Cell Bank (KCLB, Seoul, South Korea).

Techniques: Immunostaining, Marker, Control, Gene Expression, Expressing, Concentration Assay