mouse leydig tumor cell line  (ATCC)

Journal: Current Research in Toxicology

Article Title: Ethylene dimethanesulfonate effects on gene promoter activities related to the endocrine function of immortalized Leydig cell lines R2C and MA-10

doi: 10.1016/j.crtox.2023.100147

Figure Lengend Snippet: (A) Representative aspect of cells after 24 h of exposure to ethylene dimethanesulfonate (EDS) of rat R2C and mouse MA-10 Leydig cells. Note that at 5 mM, the cell confluency and morphological characteristic seem qualitatively impaired (red arrows; insert), indicating the cytotoxicity caused by EDS. (B) Cell count of R2C and MA-10 cells after 4 and 24 h of EDS exposure at concentrations of 1 or 2 mM (n = 4, each in duplicate). Values are expressed as median (interquartile ranges). Kruskal-Wallis followed by Dunn’s test (p > 0.05).

Article Snippet: Rat and mouse immortalized Leydig cell lines, R2C (ATCC, Manassas, Virginia, USA) and MA-10 (ATCC, Manassas, Virginia, USA), respectively, were used for this study from passages 40 to 52 (R2C) or 08 to 31 (MA-10).

Techniques: Cell Counting

Journal: Current Research in Toxicology

Article Title: Ethylene dimethanesulfonate effects on gene promoter activities related to the endocrine function of immortalized Leydig cell lines R2C and MA-10

doi: 10.1016/j.crtox.2023.100147

Figure Lengend Snippet: A dose–response study with reporter luciferase assays to determine the effects of EDS on (A) Star , (B) Cyp17a1 , (C) Insl3 , and (D) Gsta3 promoter activity. Rat R2C (n = 4, each in triplicate) and mouse MA-10 (n = 5, each in triplicate) Leydig cells were transfected with different promoter constructs and treated for 24 h with increasing concentrations of EDS as indicated. Luciferase reporter (Luc). Values are expressed as mean ± S.E.M. ANOVA followed by Dunnett’s test (*p < 0.05 compared to the control group).

Article Snippet: Rat and mouse immortalized Leydig cell lines, R2C (ATCC, Manassas, Virginia, USA) and MA-10 (ATCC, Manassas, Virginia, USA), respectively, were used for this study from passages 40 to 52 (R2C) or 08 to 31 (MA-10).

Techniques: Luciferase, Activity Assay, Transfection, Construct, Control

Journal: Current Research in Toxicology

Article Title: Ethylene dimethanesulfonate effects on gene promoter activities related to the endocrine function of immortalized Leydig cell lines R2C and MA-10

doi: 10.1016/j.crtox.2023.100147

Figure Lengend Snippet: EDS effects on the activity of the Insl3 and Gsta3 gene promoter after 4 and 24 h of exposure to EDS of rat R2C (1 mM of EDS, n = 4–5, each in triplicate) and mouse MA-10 (2 mM of EDS, n = 5, each in triplicate) Leydig cells. Luciferase reporter (Luc). Values are expressed as mean ± S.E.M. Welch’s t -test (*p < 0.05).

Article Snippet: Rat and mouse immortalized Leydig cell lines, R2C (ATCC, Manassas, Virginia, USA) and MA-10 (ATCC, Manassas, Virginia, USA), respectively, were used for this study from passages 40 to 52 (R2C) or 08 to 31 (MA-10).

Techniques: Activity Assay, Luciferase

Journal: Current Research in Toxicology

Article Title: Ethylene dimethanesulfonate effects on gene promoter activities related to the endocrine function of immortalized Leydig cell lines R2C and MA-10

doi: 10.1016/j.crtox.2023.100147

Figure Lengend Snippet: EDS effects on Star promoter activity after 4 h of exposure to EDS of rat R2C (1 mM of EDS, n = 5, each in triplicate) and mouse MA-10 (2 mM of EDS, n = 5, each in triplicate) Leydig cells. EDS effects on Star promoter activity after 24 h of exposure to EDS of rat R2C (1 mM of EDS, n = 3, each in triplicate) and mouse MA-10 (2 mM of EDS, n = 5, each in triplicate) Leydig cells, DC3 granulosa cells (1 mM of EDS, n = 5, each in triplicate), and MSC-1 Sertoli cells (2 mM of EDS, n = 5, each in triplicate). EDS effects were assessed in basal and stimulated (0.5 mM 8Br-cAMP) conditions in R2C Leydig cells. Luciferase reporter (Luc). Values are expressed as mean ± S.E.M. Welch’s t -test (*p < 0.05).

Article Snippet: Rat and mouse immortalized Leydig cell lines, R2C (ATCC, Manassas, Virginia, USA) and MA-10 (ATCC, Manassas, Virginia, USA), respectively, were used for this study from passages 40 to 52 (R2C) or 08 to 31 (MA-10).

Techniques: Activity Assay, Luciferase

Journal: Current Research in Toxicology

Article Title: Ethylene dimethanesulfonate effects on gene promoter activities related to the endocrine function of immortalized Leydig cell lines R2C and MA-10

doi: 10.1016/j.crtox.2023.100147

Figure Lengend Snippet: The EDS-responsive region is located between −400 and −195 bp of the Star promoter. Progressive 5′ deletion constructs of Star gene promoter were transfected in rat R2C Leydig cells and treated with 1 mM EDS for 24 h (n = 3–5, each in triplicate) in the absence or presence of 0.5 mM 8Br-cAMP for 4 h. Luciferase reporter (Luc). Values are expressed as mean ± S.E.M. Welch’s t -test (*p < 0.05).

Article Snippet: Rat and mouse immortalized Leydig cell lines, R2C (ATCC, Manassas, Virginia, USA) and MA-10 (ATCC, Manassas, Virginia, USA), respectively, were used for this study from passages 40 to 52 (R2C) or 08 to 31 (MA-10).

Techniques: Construct, Transfection, Luciferase

Journal: International journal of molecular sciences

Article Title: Highly-Sensitive In Vitro Bioassays for FSH, TSH, PTH, Kp, and OT in Addition to LH in Mouse Leydig Tumor Cell.

doi: 10.3390/ijms241512047

Figure Lengend Snippet: Figure 1. Effect of 1/10 diluted sera from various species or BSA on the cAMP response to ovine LH. (A) Effect of sera. Using the standard protocol including forskolin and oxytocin [1] in the mLTC medium, the sera from various species were present at 1% final concentration. (B) Effect of BSA (0 or 1 mg/mL) during oLH dilution (BSA dil) compared to the same final BSA quantity added separately to the cell medium (BSA cell). AUC: Area Under 60 min kinetics Curves.

Article Snippet: The mouse Leydig Tumor Cells (mLTC) were obtained from the American Tissue and Cell Collection (ATCC) (LGC Standards, Molsheim, France) and have been regularly tested negative for mycoplasma contamination all over our studies.

Techniques: Concentration Assay

Journal: International journal of molecular sciences

Article Title: Highly-Sensitive In Vitro Bioassays for FSH, TSH, PTH, Kp, and OT in Addition to LH in Mouse Leydig Tumor Cell.

doi: 10.3390/ijms241512047

Figure Lengend Snippet: Figure 2. Upper panel: Cyclic AMP response to hFSH (A) and hLH (B) after transfection of mLTC with increasing concentrations of hFSHR expression plasmid. The mLTC cells were transfected with no or 6.25 to 100 ng hFSHR plasmid before challenging with 10–1000 pM concentrations hFSH (A) or hLH (B). (C) hFSH dose–response curves when the hormone is diluted in RPMI medium alone (blue dots) or completed with 1 mg/mL BSA (red dots).

Article Snippet: The mouse Leydig Tumor Cells (mLTC) were obtained from the American Tissue and Cell Collection (ATCC) (LGC Standards, Molsheim, France) and have been regularly tested negative for mycoplasma contamination all over our studies.

Techniques: Transfection, Expressing, Plasmid Preparation

Journal: International journal of molecular sciences

Article Title: Highly-Sensitive In Vitro Bioassays for FSH, TSH, PTH, Kp, and OT in Addition to LH in Mouse Leydig Tumor Cell.

doi: 10.3390/ijms241512047

Figure Lengend Snippet: Figure 3. Cyclic AMP response to ovine TSH in mLTC transfected with the oTSHR. (A): oTSH dose–response curves in the absence or presence of 1/20 ovine serum (0.5% final concentration). (B): oTSH dose–response curves when the hormone is diluted in RPMI medium alone (blue dots) or completed with 1 mg/mL BSA (green dots). (C): oTSH concentration profiles in serums at 20-fold dilution measured by the in vitro bioassay compared to oTSH RIA. The red curve shows the AUC in the absence of TSHR (thus due only to oLH in the samples).

Article Snippet: The mouse Leydig Tumor Cells (mLTC) were obtained from the American Tissue and Cell Collection (ATCC) (LGC Standards, Molsheim, France) and have been regularly tested negative for mycoplasma contamination all over our studies.

Techniques: Transfection, Concentration Assay, In Vitro, Bioassay

Journal: International journal of molecular sciences

Article Title: Highly-Sensitive In Vitro Bioassays for FSH, TSH, PTH, Kp, and OT in Addition to LH in Mouse Leydig Tumor Cell.

doi: 10.3390/ijms241512047

Figure Lengend Snippet: Figure 4. Dose-dependent cyclic AMP response to 10−14–10−8 M hPTH in mLTC transfected with hPTHR. The mLTC cells were transfected with 0 or 1.56 to 12.5 ng PTHR plasmid per well before challenging with 10−14–10−8 M hPTH 1–34.

Article Snippet: The mouse Leydig Tumor Cells (mLTC) were obtained from the American Tissue and Cell Collection (ATCC) (LGC Standards, Molsheim, France) and have been regularly tested negative for mycoplasma contamination all over our studies.

Techniques: Transfection, Plasmid Preparation

Journal: International journal of molecular sciences

Article Title: Highly-Sensitive In Vitro Bioassays for FSH, TSH, PTH, Kp, and OT in Addition to LH in Mouse Leydig Tumor Cell.

doi: 10.3390/ijms241512047

Figure Lengend Snippet: Figure 6. Dose-dependent cyclic AMP response to 10−12–10−7 M kisspeptin variants together with 0.3 nM oLH. The mLTC were transfected with 100 ng/well of the GPR54 hKp receptor plasmid before challenging with Kp and oLH added together at t48h for luminescence recording.

Article Snippet: The mouse Leydig Tumor Cells (mLTC) were obtained from the American Tissue and Cell Collection (ATCC) (LGC Standards, Molsheim, France) and have been regularly tested negative for mycoplasma contamination all over our studies.

Techniques: Transfection, Plasmid Preparation

Journal: International journal of molecular sciences

Article Title: Highly-Sensitive In Vitro Bioassays for FSH, TSH, PTH, Kp, and OT in Addition to LH in Mouse Leydig Tumor Cell.

doi: 10.3390/ijms241512047

Figure Lengend Snippet: Figure 8. Effect of OT (A) or AVP (B) on the cyclic AMP response to hCG in mLTC. The mLTC cells were pre-incubated for 2 h with the shown concentrations of 10−20–10−6 M OT (A) or AVP (B) before stimulation with 2.5 × 10−10 M hCG and luminescence recording.

Article Snippet: The mouse Leydig Tumor Cells (mLTC) were obtained from the American Tissue and Cell Collection (ATCC) (LGC Standards, Molsheim, France) and have been regularly tested negative for mycoplasma contamination all over our studies.

Techniques: Incubation