Human hepatic HepaRG cells were seeded at low density in William’s E medium with GlutaMAX (Gibco), supplemented with 10% FBS (Hyclone II GE), 1% penicillin/streptomycin (Sigma), 5 µg/mL insulin (Sigma), 0.5 µM hydrocortisone hemisuccinate (Sigma). After 1 week at confluence, cells were shifted into the same medium supplemented with 50 µM hydrocortisone hemisuccinate and 2% DMSO (Sigma) for 2 more weeks to obtain confluent differentiated cultures. Cells were then treated with different drugs (Supplementary Fig. 1c ) for the indicated time at the following final concentrations: sodium oleate 250 μM (Sigma); S3I-201 100 μM alias NSC74859 (Selleckchem.); N-acetylcisteina 10 mM (Sigma); Ruxolitinib 1 μM (Selleckchem.). Cytoxicity of compounds was tested by MTT assay (Supplementary methods and Supplementary Fig. 1d ).
Hydrocortisone hemisuccinate
Manufactured by Merck Group
Sourced in United States, Germany
Hydrocortisone hemisuccinate is a chemical compound used in laboratory settings. It is a derivative of the natural steroid hormone hydrocortisone. The primary function of hydrocortisone hemisuccinate is to serve as a chemical reagent for research and analytical purposes.
Automatically generated - may contain errors
Lab products found in correlation
Insulin, by Merck Group (14 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (7 mentions)
William s e medium, by Thermo Fisher Scientific (7 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (6 mentions)
Streptomycin, by Merck Group (5 mentions)
Penicillin streptomycin, by Merck Group (5 mentions)
Penicillin, by Capricorn (5 mentions)
Streptomycin, by Capricorn (5 mentions)
Glutamax, by Thermo Fisher Scientific (5 mentions)
Trypsin edta, by Thermo Fisher Scientific (5 mentions)
Human insulin, by Merck Group (4 mentions)
Penicillin, by Merck Group (4 mentions)
Human insulin, by PAN Biotech (4 mentions)
Fetal bovine serum (fbs), by Merck Group (3 mentions)
Penicillin, by Thermo Fisher Scientific (3 mentions)
Streptomycin, by Thermo Fisher Scientific (3 mentions)
L glutamine, by Thermo Fisher Scientific (3 mentions)
Trypsin edta, by Euroclone (2 mentions)
Heparg human progenitor hepatic cells, by Thermo Fisher Scientific (2 mentions)
35 protocols using hydrocortisone hemisuccinate
1
Hepatic HepaRG Cell Culture and Drug Treatments
2
Culturing Human Hepatic Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HepG2 human hepatocarcinoma cells were maintained in MEM medium (Gibco, Life Technology) supplemented with 10% fetal bovine serum (Gibco, Life Technology) in the presence of 100 U/ml penicillin and 100 mg/ml streptomycin (Sigma-Aldrich, USA). HepaRG human progenitor hepatic cells (Thermo Fisher Scientific) were maintained according to the manufacturer’s recommendations. Briefly, the cells were grown in William’s E medium (Thermo Fisher Scientific) supplemented with Glutamax (Gibco), 5 μg/mL human insulin (Sigma-Aldrich) and 50 μM hydrocortisone hemisuccinate (Sigma-Aldrich) for 14 days. All cells were kept at 37°C in a humidified 5% CO2 cell culture incubator and were passaged using trypsin-EDTA (Euroclone).
3
Hepatocyte Culture Staining Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Acetaminophen (APAP), collagen, penicillin/streptomycin, Williams’ Medium E, hydrocortisone hemisuccinate, insulin, dimethyl sulfoxide (DMSO), DAPI (4′,6-Diamidino-2-phenylindole dihydrochloride), Mowiol and 6-carboxyfluorescein diacetate were purchased from Sigma-Aldrich (Steinheim, Germany); fetal bovine serum and glutamine were purchased from Biochrom (Berlin, Germany). For immunofluorescence staining, the following antibodies were used: anti-ZO1 mouse IgG (BD Biosciences, Heidelberg, Germany), anti CYP2E1 rabbit and anti CYP3A4 rabbit IgG (Bioss Antibodies, Woburn, MA, USA), AlexaFluor594 labeled goat anti-mouse and AlexaFluor488 labeled goat anti-rabbit antibody (Invitrogen, Darmstadt, Germany). A live/dead cell staining kit II was purchased from Promocell.
4
Hepatocyte Cell Line Cultivation Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Undifferentiated human HepaRG cells (HPR101) were purchased from Biopredic International (Rennes, France) and human normal immortalized THLE2 and hepatoma Huh7 cells were obtained from American Type Culture Collection. William’s E medium with no glutamine, bronchial epithelium growth medium (BEGM) medium with complete supplement, Dulbecco’s modified Eagle medium (DMEM), fetal bovine serum (FBS), fibronectin, phosphate buffered saline (PBS), L-glutamine, penicillin/streptomycin, and trypsin-EDTA were purchased from Thermo Fisher Scientific (Rockford, IL, USA). Collagen type I was purchased from Advanced Biomatrix (San Diego, CA, USA). Insulin, hydrocortisone hemisuccinate, dimethyl sulfoxide (DMSO), methanol, acetonitrile anhydrous, CHIR, 30% wt H2O2 solution, and agarose (low gelling temperature) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Chlorzoxazone, tamoxifen, isoniazid, bromobenzene, and APAP were acquired from Sigma-Aldrich (USA).
5
Inducing Differentiation of HepaRG Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HepaRG cells were purchased from Biopredic International (Saint-Grégoire, France) and cultivated at 37 °C for 14 days in William's E medium supplemented with 10 % fetal bovine serum (FBS), 5 μg/mL insulin (medium and both supplements from PAN-Biotech GmbH, Aidenbach, Germany), 50 μM hydrocortisone hemisuccinate (Sigma Aldrich, Steinheim, Germany), 100 U/mL penicillin and 100 μg/mL streptomycin (Capricorn Scientific, Ebsdorfergrund, Germany), with a change of medium every 2-3 days. After 14 days, 1.7 % dimethyl sulfoxide (DMSO) was added to the medium for additional 14 days to induce differentiation of the HepaRG cells. The medium was then changed to serum-free assay medium (SFM) for two days. SFM was adapted from Klein et al. (2014[31 (link)]). It consists of William's E medium without phenol red (PAN-Biotech GmbH, Aidenbach, Germany) supplemented with 100 U/mL penicillin and 100 μg/mL streptomycin, 2.5 μM hydrocortisone hemisuccinate, 10 ng/mL human hepatocyte growth factor (Biomol GmbH, Hamburg, Germany), 2 ng/mL mouse epidermal growth factor (Sigma Aldrich, Steinheim, Germany) and 0.5 % DMSO.
6
Hepatocyte Cell Culture Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HepG2-NTCP K7 cells and Huh-7 NTCP cells (Ko et al., 2018 (link); Meredith et al., 2016 (link)) were cultured in Dulbecco’s Modified Eagles Medium F12 supplemented with 10% foetal bovine serum (FBS) and penicillin/streptomycin. HepaRG cells were cultured in Williams E medium supplemented with 10% FBS, 50 U penicillin/streptomycin/mL, 5 μg human insulin/mL and 5 × 10−7 M hydrocortisone hemisuccinate (Sigma). Cells were seeded and expanded for 2 weeks and differentiated for another 2 weeks in the presence of 1.8% dimethyl sulfoxide (DMSO) (Gripon et al., 2002 (link)).
7
Differentiation and Treatment of HepaRG Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Undifferentiated HepaRG cells (Biopredic International, Saint Grégoire, France) were seeded in 96-well plates (9000 cells per well; for γH2AX staining), in 12-well plates (100,000 cells per well, for the Comet assay) or in 6-well plates (200,000 cells per well; for real-time RT-PCR) and cultured as previously described (Gripon et al., 2002 (link); Luckert et al., 2018 (link)). Briefly, cells were cultivated and proliferated at 37 °C in a humidified atmosphere in William's E medium with 2 mM glutamine (Pan-Biotech, Aidenbach, Germany) supplemented with 10% fetal calf serum (FCS; Pan-Biotech), 100 U/mL penicillin, and 100 μg/mL streptomycin (both from Capricorn Scientific, Ebsdorfergrund, Germany), 0.05% human insulin (Pan-Biotech), and 50 μM hydrocortisone hemisuccinate (Sigma-Aldrich). After a 2-week proliferation phase, the cells were differentiated in the abovementioned culture medium additionally supplemented with 1.7% DMSO, for two further weeks. Differentiated HepaRG cells were treated with the test compounds in differentiation medium for 24 h.
8
Cell culture and CD8+ T cell clones
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell lines of hepatic origin: Huh7_Lunet, HepG2, Huh6.1 and cell lines of non-hepatic origin: HEK 293T, HeLa, 5637, LS180, CaCo2 and A549 were cultured in Dulbeccós modified essential medium (DMEM; Life Technologies, Darmstadt, Germany) supplemented with 2 mM L-glutamine, 0.1 mM nonessential amino acids, 100 U/ml of penicillin, 100 μg/ml of streptomycin (all Life Technologies) and 10% fetal calf serum (FCS). Huh7_Lunet cells have been described previously [10] (link). Huh6.1 cells are a sub-clone of the human hepatoma cell line Huh6 and were generated by curing Huh6 cells harboring a persistent GT1b Con-1 HCV replicon with CsA and daclatasvir [11] (link), [12] (link). HepaRG cells were cultured in Williams E medium (Life Technologies) supplemented with 5 μg/ml of insulin, 50 μM hydrocortisone hemisuccinate (both Sigma Aldrich, Steinheim, Germany), 100 U/ml of penicillin, 100 U/ml of streptomycin and 10% FCS. NS5B2594-2602 (NS5BEM)-specific CD8+ T cell clones were generated by limited dilution after sorting of tet+ CD8+ T cells as previously described [9] (link). All cells were incubated in a 37 °C incubator at 95% humidity and 5% CO2. Alisporivir (Novartis, Basel, Switzerland) was dissolved to 20 mg/ml in DMSO. Detailed information on other inhibitors can be found in [13] (link) and Supplementary materials and methods .
9
Culturing Differentiated and Undifferentiated HepaRG Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Both differentiated and undifferentiated HepaRG cells were purchased from BIOPREDIC INTERNATIONAL. After thawing, the undifferentiated HepaRG cells were cultured in complete William's E medium (William's E medium (Gibco) supplemented with 10% fetal bovine serum, 1% penicillin/streptomycin/glutamine solution, 5 mg/mL insulin (Sigma-Aldrich), and 0.5 μM hydrocortisone hemisuccinate (Sigma-Aldrich)), as previously described 15 (link). The differentiated HepaRG were maintained in HepaRG™ Maintenance/Metabolism Medium (BIOPREDIC INTERNATIONAL, France) for further hepatocyte function analysis. HepG2 cell lines were obtained from ATCC and were cultured in DMEM (Gibco) containing 10% FBS and 1% penicillin-streptomycin (Gibco).
10
Culturing Human Progenitor Hepatic Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HepaRG human progenitor hepatic cells (Thermo Fisher Scientific) were seeded at a density of 10,000 cells/well and maintained at 37 °C in a humidified 5% CO2 atmosphere in William's E medium (Thermo Fisher Scientific) supplemented with 1% Glutamax (Gibco, Thermo Fisher Scientific; catalog number: 35050061), 5 μg/mL human insulin (Lonza) and 50 μM hydrocortisone hemisuccinate (Sigma-Aldrich). Cells were used between passage 2 and 10, and passages were implemented using trypsin-EDTA (Euroclone).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!