HeLa cells were from IGBMC (Strasbourg, France) with authentication and they were not used beyond passage 20 from original derivation. HeLa cells were routinely tested for mycoplasma contaminations. HeLa cells were cultured in Minimum Essential Media (MEM) (11095-080; Life technologies) supplemented with L-glutamine-penicillin-streptomycin solution (G6784; Sigma-Aldrich) and containing 10% fetal bovine serum (FBS). The medium was changed every 24 hours. Medium replenishment experiment was carried out using DMEM (11960-044; Life technologies, [high glucose, no glutamine]) supplemented with L-glutamine-penicillin-streptomycin solution (G6784; Sigma-Aldrich) and containing 10% FBS.
L glutamine penicillin streptomycin solution
Manufactured by Merck Group
Sourced in United States, Germany
L-glutamine-penicillin-streptomycin solution is a common cell culture supplement used to support cell growth and viability in vitro. It contains the amino acid L-glutamine, as well as the antibiotics penicillin and streptomycin, which help prevent bacterial contamination in cell cultures.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (11 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (5 mentions)
Fetal bovine serum (fbs), by Merck Group (4 mentions)
Rpmi 1640 medium, by Merck Group (3 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (3 mentions)
Mem nonessential amino acids solution 100x, by Thermo Fisher Scientific (3 mentions)
Trypsin edta, by Thermo Fisher Scientific (3 mentions)
Sodium pyruvate, by Merck Group (2 mentions)
Minimum essential medium (mem), by Thermo Fisher Scientific (2 mentions)
H1299, by American Type Culture Collection (2 mentions)
Rpmi 1640, by Lonza (2 mentions)
Fetal calf serum (fcs), by Merck Group (2 mentions)
Rpmi 1640, by Merck Group (2 mentions)
Penicillin streptomycin, by Merck Group (2 mentions)
Crl 2266, by American Type Culture Collection (2 mentions)
Crl 2271, by American Type Culture Collection (2 mentions)
Ccl 127, by American Type Culture Collection (2 mentions)
Dmem f12 1 1, by Thermo Fisher Scientific (2 mentions)
Ack lysing buffer, by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
40 protocols using l glutamine penicillin streptomycin solution
1
HeLa Cell Culture Protocol
2
Antiviral Activity of 1,2,3-Triazole Derivatives
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The 1,2,3-triazole derivatives were diluted at the concentration of 50 mM in dimethylsulfoxide (DMSO) and stored at -20°C. We tested ACV as a reference antiviral drug to compare our data.
These compounds were evaluated in vitro on human fibroblast (HFL1 ATCC ® CCL-153 TM ) cells which were cultivated at 37°C and 5% CO 2, with Dulbecco's Modification of Eagle's medium (DMEM), with 4.5 g/l glucose, l-glutamine and sodium pyruvate (Sigma-Aldrich, Saint Louis, MO, USA), supplemented with 10% fetal bovine serum (Gibco, Carlsbad, CA, USA), and 1% of l-glutamine-penicillin-streptomycin solution (Sigma-Aldrich).
To perform the plaque assay, we used rabbit skin (RS) cells cultivated at 37°C and 5% CO 2 with Minimum Essential Medium Eagle (MEM), with Earle's salts and l-glutamine (Sigma-Aldrich), supplemented with 5% bovine serum (Gibco) and 1% of l-glutamine-penicillin-streptomycin solution (Sigma-Aldrich).
The cells were cultivated until they achieved around 90% of confluence when they were transferred to plates of 24 or 96 wells depending on the assay.
HSV-1 strain 17syn+ was used for the experiments. We also tested the compounds on the ACV-resistant clinical strain HO-1 (kindly provided by the D Phelan, College of Medicine, University of Florida, personal communication). Virus stock cultures were prepared from supernatants of infected cells and stored at -80°C until use.
These compounds were evaluated in vitro on human fibroblast (HFL1 ATCC ® CCL-153 TM ) cells which were cultivated at 37°C and 5% CO 2, with Dulbecco's Modification of Eagle's medium (DMEM), with 4.5 g/l glucose, l-glutamine and sodium pyruvate (Sigma-Aldrich, Saint Louis, MO, USA), supplemented with 10% fetal bovine serum (Gibco, Carlsbad, CA, USA), and 1% of l-glutamine-penicillin-streptomycin solution (Sigma-Aldrich).
To perform the plaque assay, we used rabbit skin (RS) cells cultivated at 37°C and 5% CO 2 with Minimum Essential Medium Eagle (MEM), with Earle's salts and l-glutamine (Sigma-Aldrich), supplemented with 5% bovine serum (Gibco) and 1% of l-glutamine-penicillin-streptomycin solution (Sigma-Aldrich).
The cells were cultivated until they achieved around 90% of confluence when they were transferred to plates of 24 or 96 wells depending on the assay.
HSV-1 strain 17syn+ was used for the experiments. We also tested the compounds on the ACV-resistant clinical strain HO-1 (kindly provided by the D Phelan, College of Medicine, University of Florida, personal communication). Virus stock cultures were prepared from supernatants of infected cells and stored at -80°C until use.
3
HeLa Cell Culture Protocol
4
Cultivation of UT-7 and C6/36 Cell Lines
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UT-7 cells (human megakaryocytic cell line) were obtained from DSMZ-German Collection of Microorganisms and Cell Cultures (Braunschweig, Germany; Catalog number: ACC-137). Cells were grown according to manufacturer’s instructions. Briefly, following revival from liquid nitrogen, cells were cultured in MEMα–no nucleosides (Gibco) containing 40% FBS and 5ng/mL GM-CSF (Peprotech) for 6 days, after which FBS was reduced to 20%. Cells were split every three days, and replated at 5x105 cells/mL in fresh media. Cells were incubated at 37°C at 5% CO2 in a humidified incubator. All UT-7 cells used for experiments were below passage 25.
C6/36 cells (Aedes albopictus cell line) were obtained from ATCC (CRL-1600). Cells were cultured in Minimum Essential Media (HyClone) containing 10% FBS, 1% Sodium Pyruvate (Sigma), 1% MEM Non-Essential Amino Acids (Sigma), and 1% Penicillin/Streptomycin/L-Glutamine solution (Sigma). Cells were incubated at 29°C at 5% CO2 in a humidified incubator.
C6/36 cells (Aedes albopictus cell line) were obtained from ATCC (CRL-1600). Cells were cultured in Minimum Essential Media (HyClone) containing 10% FBS, 1% Sodium Pyruvate (Sigma), 1% MEM Non-Essential Amino Acids (Sigma), and 1% Penicillin/Streptomycin/L-Glutamine solution (Sigma). Cells were incubated at 29°C at 5% CO2 in a humidified incubator.
5
Uterine Weights and Immune Cell Isolation
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Uterine wet weights were recorded. Bone marrow (BM) cells were harvested by flushing the cavity of one femur and one humerus with PBS. Lymph nodes draining the joints (subiliac, popliteal, sciatic, proper and accessory axillary) were dissected and mashed through a 70 μm nylon mesh filter and re-suspended in complete medium [phenol red-free RPMI 1640 (PAA Laboratories, Pasching, Austria) supplemented with 10% dextran-coated charcoal hormone-stripped FCS (Sigma) and 1% penicillin-streptomycin-l -glutamine solution (Sigma)]. Erythrocytes in BM were lysed by using Tris-buffered 0.83% NH4Cl solution. Cells were counted using an automated cell counter (Sysmex Europe, Nordenstedt, Germany).
6
Cytotoxic Activity of Quercetin on Neuroblastoma and Fibroblast Cells
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In the cytotoxic activity study, SH-SY5Y (neuroblastoma) cell line (ATCC CRL 2266) and NIH 3T3 ( broblast) cell line (ATCC CRL-1658) was obtained from American Type Culture Collection. Dulbecco's modi ed Eagle's medium (DMEM), fetal bovine serum (FBS), and Quercetin were purchased from Merck Millipore. Chitosan (400 kDa, DD 92), phosphate buffer saline (PBS), and polyvinyl pyrrolidone (PVP) were obtained from Sigma-Aldrich. Penicillin-Streptomycin-L-glutamine solution was purchased from Sigma-Aldrich. XTT reagent (Roche Diagnostic) was utilized in cytotoxic activity studies. SH-SY5Y and NIH 3T3 cells were seeded in DMEM containing FBS (10 %), penicillin (100 IU/mL), L-glutamine (1 %), and streptomycin (10 mg/mL). Well plates including cells were incubated in an incubator at 5 % CO 2 and 37 °C. The cytotoxic activity studies were performed when cells reached at least 80 % con uence [25] .
7
Cell Culture Protocols for NSCLC and Immune Cells
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NSCLC cell lines (A549, H1299, H460), Lewis lung cancer (LLC) cell line, and 293 T cell line were obtained from the American Type Culture Collection (ATCC, USA) and validated by short-tandem-repeat (STR) analysis (except for LLC). Cells were cultured in either RPMI-1640 (for NSCLC cell lines) or DMEM (for LLC cells and 293 T cells) containing 10% fetal bovine serum and maintained at 37 °C in a humidified 5% CO2 incubator. Peripheral blood mononuclear cells (PBMCs) were cultured in T cell medium (RPMI-1640 supplemented with 10% human serum, 5% L-glutamine-penicillin-streptomycin solution (Sigma-Aldrich, USA), and IL-2 (100 IU/mL).
8
Epirubicin and Inflammasome Activation in THP-1 Cells
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Cells were cultivated in RPMI-1640 medium (Biochrom AG, Berlin, Germany) supplemented with 10% fetal bovine serum (FBS; Pan Biotech, Aidenbach, Germany) and 1% L-glutamine-penicillin-streptomycin solution (GPS; Sigma-Aldrich, St. Louis, Missouri, USA). Prior to further stimulation, THP-1 were differentiated with 100 ng/mL phorbol 12-myristate 13-acetate (PMA; Abcam, Cambridge, UK) for 18 h.
Cells were treated with freshly prepared, sterile-filtered epirubicin (Sigma Aldrich) at concentrations between 0.01 to 5 µg/mL for 24 h, unless stated otherwise. After epirubicin treatment, PM and THP-1 were stimulated for 3 h with LPS from E.coli K12 (InvivoGen, San Diego, CA, USA) at concentrations of 10 or 100 ng/mL, respectively, followed by treatment with freshly prepared 1 mM ATP (InvivoGen, San Diego, CA, USA) for 1 h. For the inhibition of autophagy, 300 nM bafilomycin (Invivogen, San Diego, CA, USA) or 10 mM 3-methyl adenine (3-MA; Invivogen, San Diego, CA, USA) were added to the cells simultaneously with LPS, and maintained until the end of the experiment. For TLR2 ligation, cells were incubated either with 108 cells/mL of heat-killed listeria monocytogenes (HKLM; Invivogen, San Diego, CA, USA) or 100 ng/mL Pam3CSK4 (Invivogen, San Diego, CA, USA) for 4 h.
Cells were treated with freshly prepared, sterile-filtered epirubicin (Sigma Aldrich) at concentrations between 0.01 to 5 µg/mL for 24 h, unless stated otherwise. After epirubicin treatment, PM and THP-1 were stimulated for 3 h with LPS from E.coli K12 (InvivoGen, San Diego, CA, USA) at concentrations of 10 or 100 ng/mL, respectively, followed by treatment with freshly prepared 1 mM ATP (InvivoGen, San Diego, CA, USA) for 1 h. For the inhibition of autophagy, 300 nM bafilomycin (Invivogen, San Diego, CA, USA) or 10 mM 3-methyl adenine (3-MA; Invivogen, San Diego, CA, USA) were added to the cells simultaneously with LPS, and maintained until the end of the experiment. For TLR2 ligation, cells were incubated either with 108 cells/mL of heat-killed listeria monocytogenes (HKLM; Invivogen, San Diego, CA, USA) or 100 ng/mL Pam3CSK4 (Invivogen, San Diego, CA, USA) for 4 h.
9
Isolation and Culture of Rat Islet Cells
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Animal studies were approved by the Geneva Institutional Animal Care and Use Committee. Islets were isolated from male Sprague Dawley rats (350 g). Ten milligrams of collagenase (Sigma Aldrich, St. Louis, MO, USA) were resuspended in 10 mL of Hanks’ balanced salt solution (Bichsel, Interlaken, Germany) and were injected into the pancreas via the common bile duct of euthanized rats. The pancreas was excised and digested 10 min at 37°C. Islets were purified by Ficoll density gradient separation and dissociated into cells by an incubation of 4 min at 37°C in a solution of 0.05% trypsin supplemented with 0.48 mM ethylenediaminetetraacetic acid (Gibco, New York, NY, USA). Rat islet cells were incubated at a concentration of 100 000 cells/10 mL in 10 cm-nonadherent petri dishes for 24 h at 37°C with 5% CO2 using Dulbecco’s modified Eagle medium (DMEM; Gibco) supplemented with L-glutamine-penicillin-streptomycin solution (Sigma), fetal calf serum (FCS; Biochrom, Berlin, Germany), sodium pyruvate (Sigma; hereafter referred as complete DMEM) and 11.2 mM glucose.
10
Cell Viability Assay of MDA-MB-231 and L929 Cells
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Human breast cancer cell line MDA-MB-231 (HTB-26) and mouse subcutaneous connective tissue cell line L929 (CRL-6364) were obtained from ATCC. Dulbecco’s modified Eagle’s medium, fetal bovine serum and phosphate buffer saline were supplied from PAA Ltd. (France). Trypsin-EDTA was purchased from Biological Industries Ltd. (Haemek, Israel). l -glutamine–penicillin–streptomycin solution was bought from Sigma-Aldrich. XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) reagent was purchased from Roche Diagnostic. Both cell lines were maintained in DMEM containing 10 % FBS, 1 % l -glutamine, 100 IU/mL penicillin, and 10 mg/mL streptomycin and cultured in a humidified atmosphere with 5 % CO2 at 37 °C. The cells were used for the experiments when they reached 85–90 % confluence.
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