human normal liver cell line wrl 68  (ATCC)

Journal: Advanced Science

Article Title: A Single‐Cell Metabolic Profiling Characterizes Human Aging via SlipChip‐SERS

doi: 10.1002/advs.202406668

Figure Lengend Snippet: Spermine promotes cellular senescence in vitro. a) Quantification of the nuclear size of GM00038 cells treated with spermine for 14 days. Number of cells per group > 1500, cells from three biological replicates. b) The viability of GM00038 cells was examined by CCK8 after being treated with spermine (1 µg mL −1 ). n = 3 biological replicates. c) Cell cycle analysis of GM00038 cells treated with spermine (1 µg mL −1 ) for 7 days. d) mRNA levels of P16, P21, LMNB1, IL‐6, IL‐8, and IL‐1A in GM00038 cells treated with spermine (1 µg mL −1 ), normalized to GAPDH mRNA (n = 3 per group). e) Representative images of SA‐β‐gal (n = 3 per group/4 images per n), EdU (yellow; n = 3 per group/4 images per n), and DAPI labeled nuclei (blue) of HUVECs treated with spermine (2 µg mL −1 ) for 5 days. Scale bar, 100 µm. f,g) Quantification of SA‐β‐gal + cells (f) and EdU + cells (g) in (d). At least 100 cells per n were calculated. h) mRNA levels of P16, P21, LMNB1, IL‐6, IL‐8, and IL‐1A in HUVECs treated with spermine (2 µg mL −1 ), normalized to GAPDH mRNA. (n = 3 per group). i) Representative images of SA‐β‐gal (n = 3 per group/4 images per n), EdU (yellow; n = 3 per group/4 images per n), and DAPI labeled nuclei (blue) of WRL68 treated with spermine (2 µg mL −1 ) for 12 days. Scale bar, 100 µm. j,k) Quantification of SA‐β‐gal + cells (j) and EdU + cells (k) in (h). At least 800 cells per n were calculated. l) mRNA levels of P16, P21, LMNB1, IL‐6, IL‐8, and IL‐1A in WRL68 treated with spermine (2 µg mL −1 ) for 7 days, normalized to GAPDH mRNA. (n = 3 per group). m) H 2 O 2 levels in GM00038 treated with spermine (1 µg mL −1 ) for 14 days (left, n = 3 per group). The amount of H 2 O 2 released into the cell culture medium within 24 h after GM00038 cells were treated with spermine (1 µg mL −1 ) for 13 days (right, n = 3 per group). n) mRNA levels of SMS, SMOX and SSAT in GM00038 cells treated with spermine (1 µg mL −1 ) for 14 days, normalized to GAPDH mRNA (n = 5 per group). o) Exogenous Spermine treatment could induce typical cellular senescence phenotype and lead to increased overall H 2 O 2 level in vitro. Data are means ± SD of biologically independent samples. Statistical significance was calculated using a two‐tailed t ‐test (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).

Article Snippet: Human normal liver cells (WRL‐68) were obtained from ATCC (United States).

Techniques: In Vitro, Cell Cycle Assay, Labeling, Cell Culture, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Protective Effect of Antrodia cinnamomea Extract against Irradiation-Induced Acute Hepatitis

doi: 10.3390/ijms20040846

Figure Lengend Snippet: The protective effects of Antrodia cinnamomea extract (ACE) against irradiation in human normal liver CL48 cells. ( A ) Cytotoxicity of irradiation in CL48. Cells were treated with different dosages (0, 5, 10, 15, and 20 Gy) of irradiation and counted by hemocytometer at intervals as indicated. ( B ) Determination of the optimal ACE dosages. Cells were treated with different concentrations of ACE for 24 h or 48 h, followed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay for cell survival. For the control group, the same concentrations of ethanol were used for each ACE treatment experiment. ( C ) The protective effect of ACE against irradiation. Cells were pre-treated with different concentrations of ACE for 16 h followed by various dosages of irradiation; they were evaluated by MTT assay 48 h later. Cell survival was evaluated by MTT assay. Results were obtained from three independent experiments; each experiment was done in triplicate. **, p < 0.01, as compared with the control group.

Article Snippet: Normal human embryonic liver cell line CL48 cells (American Type Culture Collection) were cultured in high glucose DMEM with 10% fetal bovine serum (FBS), 0.1 mg/mL of streptomycin, and 100 units/mL of penicillin, at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Irradiation, MTT Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: Protective Effect of Antrodia cinnamomea Extract against Irradiation-Induced Acute Hepatitis

doi: 10.3390/ijms20040846

Figure Lengend Snippet: Protective effects of ACE against irradiation-induced apoptosis. CL48 cells were pre-treated with various concentrations of ACE for 16 h followed by irradiation at 15 Gy. Cells were harvested 48 h later for determination of caspase-3 activity ( A ), caspase-8 activity ( B ), and caspase-9 activity ( C ). The fold differences of the three caspase activities relative to individual control groups were also presented ( D ). For apoptosis analysis, cells were harvested 30 h after treatment followed by staining with PI and Annexin V ( E ). Results are representative of three independent experiments. The statistical analysis of the percentages of early plus late apoptotic cells ( F ). *, p < 0.05; **, p < 0.01, as compared with the irradiated ACE 0 μg/mL control group.

Article Snippet: Normal human embryonic liver cell line CL48 cells (American Type Culture Collection) were cultured in high glucose DMEM with 10% fetal bovine serum (FBS), 0.1 mg/mL of streptomycin, and 100 units/mL of penicillin, at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Irradiation, Activity Assay, Control, Staining

Journal: International Journal of Molecular Sciences

Article Title: Protective Effect of Antrodia cinnamomea Extract against Irradiation-Induced Acute Hepatitis

doi: 10.3390/ijms20040846

Figure Lengend Snippet: Reactive oxygen species (ROS) scavenging activity of ACE in CL48 cells. After being treated and harvested as described in the Materials and Methods section, cells were analyzed by flowcytometry after incubation with 2′,7′-dichlorofluorescin diacetate (DCFH-DA) for 15 min. ( A ) Control cells without treatment, ( B ) IR treated only, ( C ) H 2 O 2 treated only, ( D ) pre-treated with NAC before IR, ( E ) pre-treated with ACE 40 μg/mL before IR, ( F ) pre-treated with ACE 80 μg/mL before IR. ( G ) Histograms of B, E, and F were combined to show the dose-dependent ROS scavenging activity of ACE. ( H ) Fold change of control fluorescence intensity by 40 μg/mL and 80 μg/mL of ACE, respectively.

Article Snippet: Normal human embryonic liver cell line CL48 cells (American Type Culture Collection) were cultured in high glucose DMEM with 10% fetal bovine serum (FBS), 0.1 mg/mL of streptomycin, and 100 units/mL of penicillin, at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Activity Assay, Incubation, Control, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Protective Effect of Antrodia cinnamomea Extract against Irradiation-Induced Acute Hepatitis

doi: 10.3390/ijms20040846

Figure Lengend Snippet: Redox-related enzymes expression and activity profiles of CL48 hepatocytes after IR and/or ACE treatments. With or without ACE pre-treatment for 16 h, cells were irradiated with a dose of 15 Gy and harvested 24 h later for RT-PCR ( A ), Western blot ( B ), total superoxide dismutase (SOD) activity ( C ), and glutathione peroxidase (GPx) activity ( D ) analyses. Results were obtained from three independent experiments, each experiment was done in triplicate. **, p < 0.01, as compared with the control IR only group.

Article Snippet: Normal human embryonic liver cell line CL48 cells (American Type Culture Collection) were cultured in high glucose DMEM with 10% fetal bovine serum (FBS), 0.1 mg/mL of streptomycin, and 100 units/mL of penicillin, at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Expressing, Activity Assay, Irradiation, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Protective Effect of Antrodia cinnamomea Extract against Irradiation-Induced Acute Hepatitis

doi: 10.3390/ijms20040846

Figure Lengend Snippet: Enhancement of the irradiation-induced nuclear factor erythroid-2-related factor (Nrf2) expression and nuclear translocation by ACE treatment. With or without ACE pre-treatment for 16 h, CL48 cells were irradiated with a dose of 15 Gy and harvested 24 h later for Western blot ( A ) and immunofluorescence staining ( B , left) of Nrf2. The slides were counterstained with 4′,6-diamidino-2-phenylindole (DAPI) (300 nM in PBS) ( B , middle) and merged images ( B , right). ( C ) The nuclear Nrf2 positivity rates were plotted. Results were obtained from random fields of three independent experiments for each group. *, p < 0.05; **, p < 0.01, as compared with the IR only group.

Article Snippet: Normal human embryonic liver cell line CL48 cells (American Type Culture Collection) were cultured in high glucose DMEM with 10% fetal bovine serum (FBS), 0.1 mg/mL of streptomycin, and 100 units/mL of penicillin, at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Irradiation, Expressing, Translocation Assay, Western Blot, Immunofluorescence, Staining