CMT 167 cells (kind gift from Prof. Paola Allavena, Humanitas Research Hospital) were cultured in Gibco Dulbecco's Modified Eagle Medium (DMEM), high glucose (Sigma‐Aldrich, #D5796), with 10% Fetal Bovine Serum (FBS, Sigma‐Aldrich #F7524), 2 mM L‐glutamine and 100 U/mL penicillin–streptomycin (Pen/strep). 4T1 cells (CRL‐2539, American Type Culture Collection (ATCC)) were grown in RPMI 1640 Medium with 10% FBS and 100 U/mL Pen/strep. KPCY cells (2838c3, Kerafast) were cultured in DMEM (Sigma‐Aldrich #D5796) with 10% FBS and 100 U/mL Pen/strep. MC38 cells (ENH204, Kerafast) were cultured in low glucose DMEM (Sigma‐Aldrich #D6046) with 10% FBS, 2 mM L‐ glutamine, 1* non‐essential amino acids, 1 mM sodium pyruvate, 10 mM HEPES (4‐(2‐hydroxyethyl)‐1‐piperazineethanesulfonic acid), and 100 U/mL Pen/strep. All chemicals were from Sigma‐Aldrich unless specified. Cells were thawed and maintained in exponential growth for 2–3 weeks at 37°C and 5% CO2 before implantation.
D5796
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom, Poland
D5796 is a laboratory equipment product manufactured by Merck Group. It is designed for specific laboratory applications, but a detailed description is not available while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (9 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (7 mentions)
F7524, by Merck Group (6 mentions)
F2442, by Merck Group (5 mentions)
Fetal bovine serum (fbs), by Merck Group (5 mentions)
P4333, by Merck Group (4 mentions)
D6046, by Merck Group (3 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (3 mentions)
Penicillin streptomycin, by Merck Group (3 mentions)
Horse serum, by Thermo Fisher Scientific (3 mentions)
M7145, by Merck Group (3 mentions)
Collagenase type 2, by Worthington (3 mentions)
Streptomycin, by Merck Group (3 mentions)
Penicillin, by Merck Group (3 mentions)
Penicillin, by Thermo Fisher Scientific (3 mentions)
Rpmi 1640, by Merck Group (3 mentions)
Pen strep, by Thermo Fisher Scientific (2 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (2 mentions)
P0781, by Merck Group (2 mentions)
Penicillin streptomycin, by Fujifilm (2 mentions)
110 protocols using d5796
1
Cell Culture Protocols for Tumor Cell Lines
2
Cell Culture Conditions for Murine and Human Fibroblasts
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Wild type (Wt) and XPG-depleted (Xpg-/-) murine embryonic fibroblasts (MEFs) [23 (link)] were grown in Dulbecco’s Modified Eagle’s Medium (DMEM) (Sigma, D5796), supplemented with 1% penicillin/streptomycin (Gibco, 11548876) and 20% heat-inactivated fetal bovine serum (Corning, 35-015-CV). MRC5-SV, XPCS1RO-SV [24 (link)], GM14930-SV (Coriell, GM14930) and GM14931-SV (Coriell, GM14931) were grown in DMEM (Sigma, D5796), supplemented with 1% penicillin/streptomycin (Gibco, 11548876) and 10% non-heat-inactivated fetal bovine serum (Corning, 35-015-CV).). All these cell lines were grown at 37°C with 5% CO2 and 3% O2 to reduce oxidative damage. XPCS1RO-SV+XPG-GFP [25 (link)] were grown with 200 μg/mL of G418 (Sigma Aldrich, G8168-50ML). They were grown at 37°C with 5% CO2.
3
Glioma and Neural Progenitor Cell Cultures
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TS603, TS667, TS543 were obtained from Memorial Sloan Kettering Cancer Center (MSKCC). SU-AO3, NCH612 glioma lines were described previously [23 (link)]. The patient-derived IDH1-mutant glioma lines were routinely subjected to panel sequencing analysis [26 (link)] and immunoblotting to confirm the expression of mutant IDH1 (R132H). Glioma lines were maintained as neurospheres in NeuroCult (StemCell Technologies) with 2 µg/ml heparin sulfate (Sigma), 20 ng/ml of EGF (StemCell Technologies), and FGF2 (StemCell Technologies). PC12 cells (provided by Dr. Agarwal, Heidelberg University; Dr. Chang, Gachon University) were maintained in DMEM (D5796, Sigma) supplemented with 10% horse serum (H1138, Sigma) and 1% penicillin–streptomycin. Human neural progenitor cells (SCC008, Sigma) were maintained in DMEM (D5796, Sigma) supplemented with 10% fetal bovine serum (Gibco) and 1% penicillin–streptomycin. All cells were grown in a 5% CO2 humidified incubator at 37 °C.
4
Internalization of MSC-EVs by Endothelial Cells
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MCEC cells were seeded on glass bottom dishes (Ø 35 mm, WillCo Wells B.V.) at a density of 5 × 104 cells/dish in complete DMEM medium (#D5796, Merck) supplemented with 5% FBS (#F2442, Merck). After 24 h post seeding, cells were washed with PBS (w/o Ca2+, Mg2+, HyClone) and divided into two groups:
Control—further culture of MCECs in complete DMEM medium with 5% FBS for the following 24 h;
Microenvironment mimicking inflammation and hypoxia—further culture of MCECs in complete DMEM medium with 1% FBS, 10 ng/mL IL-1β (#211-11B, Peprotech) and 10 ng/mL TNF-α (#315-01, Peprotech) for the following 24 h.
5
Cell Proliferation and Differentiation
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DMEM (Merck #D5796) containing 10% fetal calf serum (Sigma Aldrich #F7524-500ML) and 1% Penicilin-Streptamycin-Glutamine (PSG) was used to keep cells in proliferation. Transient transfections of cell were always performed using Lipofectamine 3000 (ThermoFisher #L3000075) according to the manufacturer’s protocol for 24 h or 48 h. For differentiation of C2-cells DMEM containing 2% HS and 1% PSG was used.
6
Cell Culture and BH3 Mimetics Protocol
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U87 glioblastoma cells were purchased from the ATCC (Manassas, Virginia, USA) and cultivated in Dulbecco's modified Eagle's high glucose medium (DMEM, D5796, Merck, Darmstadt, Germany) supplemented with 10% FBS (D7524, Merck), 1% non-essential amino acids and 1% antibiotics (penicillinstreptomycin, P4333, Merck). Mantle cell lymphoma (MCL) cell line HBL-2 was obtained from Prof. Pavel Klener (HBL-2 cells were originally obtained from Prof. Martin Dreyling, Munich, Germany), and cultivated in Iscove's Modified Dulbecco's Medium media (IMDM, I7633, Merck) supplemented with 15% FBS (F7524, Merck) and 1% antibiotics (Penicillin-Streptomycin, P4333, Merck). Cultured cells were regularly tested for mycoplasma infection using the MycoAlert™ kit (LT07-318, Lonza, Basel, Switzerland), and only mycoplasma-negative cells were used. BH3 mimetics were purchased from MedChemExpress (Monmouth Junction, New Jersey, USA) and, unless stated otherwise, all other chemicals were from Merck. HCT-116 Bax/Bak -/-cells were kindly provided by Richard Youle (National Institutes of Health, Bethesda, Maryland, USA).
7
Culture of Human Liver Stem Cells
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8
Primary Neuron Culture from Mouse Cortex
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Embryonic C57BL/6J mice (15.5-16.5 days old) were used for primary neuron culture. The frontal cortices and hippocampi were dissected from the brains in cold HEPES-buffered, calcium and magnesium-free Hank's balanced salt solution (HBSS; 14175-103, Thermo Fisher Scientific, Waltham, MA, USA) and dissociated in HBSS containing 0.25% papain (LS003127, Worthington Biochemical Corporation, Lakewood, NJ, USA), 0.1% L-cysteine (C7352, Merck, Darmstadt, Germany), 0.1% bovine serum albumin (BSA; A2153, Merck), and 0.01% DNase I (D4263, Merck) at 37°C for 12 minutes. The enzymatic digestion was stopped by adding a plating medium (Dulbecco's modified Eagle medium, D5796, Merck, containing 10% fetal bovine serum). Tissues were dissociated by pipetting, centrifuged at room temperature for 10 minutes, and resuspended in the plating medium. Cells were counted, seeded at a density of 25,000 cells/cm 2 on culture plates or glass coverslips previously coated with 0.01% poly-L-lysine (P8920, Merck), and incubated at 37°C. After 2 hours, the plating medium was replaced by a culture medium (Neurobasal™ medium
9
GFP-Tagged BoHV-1 Infection in MDBK Cells
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Wild type Madin-Darby bovine kidney cells (MDBK), Cas9+/+ MDBK cells or dCas9+/+ cells [49 (link)] were cultured in DMEM (D5796, Sigma, Tokyo, Japan) supplemented with 5% horse serum (26050088, Gibco), 1% Pen/Strep (15140122, Gibco), 1% L-glutamine (25030024, Gibco), 1% NEAA (11140035, Gibco, Paisley, Scotland), and 1% sodium pyruvate (11360039, Gibco). Cells were kept in an incubator set at 37 °C with 5% CO2. A GFP tagged BoHV-1 strain based on strain Jura [53 (link)] was used throughout the experiment. The GFP protein is fused to the VP26, a minor capsid protein of BoHV-1.
10
FANCD2 and UHRF1 Localization Dynamics
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HeLa cells (female) were grown in DMEM (D5796, Sigma) supplemented with 10% FBS. EGFP-fused FANCD2 and mCherry-fused UHRF1 cDNA were expressed using a derivative of the pOZ-N plasmid and sorted using α-IL2R coupled magnetic beads (Dynabeads goat anti-mouse IgG) as described in (Nakatani and Ogryzko, 2003 (link)). Transfections of plasmid DNA were carried out using FuGENE6 (Promega) according to the manufacturer’s instructions.
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