The 293-EBNA, 293FT, U937, THP-1, HTC-116, HT-29 cell lines were obtained from ATCC (Manassas, VA, USA). NHDF (normal human dermal fibroblasts) cells were obtained from LONZA (Basel, Switzerland). U937, THP-1, HTC-116, HT-29 cells were cultured in RPMI medium (LONZA, Basel, Switzerland) with 10% fetal bovine serum; 293-EBNA in Dulbecco’s modified Eagle medium (LONZA, Basel, Switzerland) with 10% fetal bovine serum containing 250 μg/ml G418; NHDF and 293FT cells were cultured in Dulbecco’s modified Eagle medium (LONZA, Basel, Switzerland) with 10% fetal bovine serum. All the cells were maintained at 37 ̊C under a humidified atmosphere containing 5% CO2 and verified to be free of mycoplasma contamination using the MycoAlert™ Mycoplasma Detection kit (LT07–318, LONZA).
Dulbecco s modified eagle medium
Manufactured by Lonza
Sourced in Switzerland, United States, Belgium, United Kingdom
Dulbecco's Modified Eagle Medium (DMEM) is a cell culture medium formulation that provides nutrients and growth factors necessary to support the in vitro cultivation of various cell types. It is a widely used and versatile medium designed to maintain the optimal growth and proliferation of cells in laboratory settings.
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Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (19 mentions)
Fetal bovine serum (fbs), by Lonza (14 mentions)
Streptomycin, by Lonza (10 mentions)
L glutamine, by Lonza (9 mentions)
Penicillin streptomycin, by Lonza (8 mentions)
Penicillin, by Lonza (8 mentions)
Penicillin streptomycin, by Merck Group (7 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (5 mentions)
Trypsin, by Lonza (5 mentions)
L glutamine, by Merck Group (4 mentions)
Trypsin edta, by Thermo Fisher Scientific (4 mentions)
Fetal calf serum (fcs), by Lonza (4 mentions)
Rpmi medium, by Lonza (3 mentions)
Mcf 7, by American Type Culture Collection (3 mentions)
Gentamycin, by Lonza (3 mentions)
Amphotericin b, by Lonza (3 mentions)
Streptomycin, by Thermo Fisher Scientific (3 mentions)
Penicillin, by Thermo Fisher Scientific (3 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (3 mentions)
Dulbecco modified eagle medium (dmem), by Lonza (3 mentions)
94 protocols using dulbecco s modified eagle medium
1
Cell Culture Conditions for Diverse Cell Lines
2
Cultivation of Human Cell Lines
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U2OS17 cells were cultured at 37 °C in DMEM (Dulbecco’s Modified Eagle Medium 4.5 g/l glucose; Lonza,) supplemented with 10% fetal bovine serum (Lonza),
U2OS osteosarcoma and A375 melanoma cells were cultured at 37 °C in DMEM (Dulbecco’s Modified Eagle Medium; Lonza) supplemented with 10% fetal bovine serum (Lonza), 4 mM L-Glutamine (Sigma-Aldrich) and 1% antibiotic (Sigma-Aldrich).
Hker E6SFM keratinocyte cells were cultured at 37 °C in Keratinocyte-SFM Medium with L-Glutamine, EGF and BPE (Thermo Fisher Scientific) supplemented with 2 mM L-Glutamine (Sigma-Aldrich) and 1% antibiotic (Sigma-Aldrich).
U2OS and A375 cell line was purchased from ATCC, U2OS17 cells were provided by Frederic Coin24 (link), while Hker E6SFM cells were provided by Vilmos Tubak and were generated as described elsewhere in accordance with the relevant guidelines59 ,60 (link). All experimental protocols were approved by the guidelines of the University of Szeged and the Medical Research Council.
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U2OS and A375 cell line was purchased from ATCC, U2OS17 cells were provided by Frederic Coin24 (link), while Hker E6SFM cells were provided by Vilmos Tubak and were generated as described elsewhere in accordance with the relevant guidelines59 ,60 (link). All experimental protocols were approved by the guidelines of the University of Szeged and the Medical Research Council.
3
Cell Culture Conditions for Neuroblasts and Cancer Cells
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Neuro-2A cells (DMSZ ACC 148)—mouse neuroblasts with neuronal and amoeboid stem cell morphology isolated from brain tissue—were cultivated in low-glucose (1.0 g/L) Dulbecco’s Modified Eagle Medium (Lonza, Walkersville, MD, USA) supplemented with 10% FBS (Bio&SELL, Feucht/Nürnberg, Germany); 2 mM of alanyl-L-glutamine (UltraGlutamineTM I Supplement, Lonza, Walkersville, MD, USA); and 1% non-essential amino acids (Sigma-Aldrich, St. Louis, USA). HCT116 cells (ATCC® CCL-247TM), a human colon cancer cell line, were cultured in McCoy’s 5A Medium (Life Technologies, Carlsbad, CA, USA) supplemented with 10% FBS and 2 mM of glutamine. A2058 cells (ATCC® CRL-11147 TM), a human amelanotic melanoma cell line, were cultivated in high-glucose (4.5 g/L) Dulbecco’s Modified Eagle Medium (Lonza, Walkersville, MD, USA) supplemented with 10% FBS, 2 mM of glutamine and 1% non-essential amino acids. Cells were grown in humidified incubators at 5% CO2 and 37 °C.
4
Neuroblast Cell Viability on Carbon Matrices
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Three groups of samples were prepared: a polymer multiwell base, a flame retardant (PCB support) carbon matrix on a flame-retardant substrate, and a pre-adsorbed glutamate carbon matrix (glu-carbon). IMR-32 neuroblast cells from ATCC (Manassas, VA, USA) were then seeded on a multiwell plate at around 250,000 cells/cm2 and incubated in Dulbecco's Modified Eagle Medium (Lonza, Basel, Switzerland), with 200 mM of 1% L-Glutamine (Lonza, Basel, Switzerland), 10% FBS (Lonza, Basel, Switzerland), 1% penicillin/streptomycin/amphotericin B ((Lonza, Basel, Switzerland)), 1% MEM vitamin solution, (Sigma Aldrich, Darmstaadt, Germany), and 1% non-essential amino acids 100X (Sigma Aldrich, Darmstaadt, Germany), along with the three sample groups. A CellTiter-Blue assay (Promega, Madison, WI, USA) was also performed on all samples. Cell viability measurements were carried out at 24, 48, and 96 h after seeding. The control measurement of the multiwell base seeded with the cells was performed 24 h after seeding. The viability of the other substrates was normalized to the viability of the control. Three repetitive samples were applied for each group's treatment.
5
Neuroblastoma Cell Culture Protocol
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Dulbecco’s modified eagle medium (DMEM), Ham’s F-12 medium (F12), streptomycin, L-glutamine, fetal bovine serum (FBS) were provided by Lonza (Verviers, Belgium). Human neuroblastoma (SH-SY5Y line) cells were cultivated not longer than 20 passages in full medium, i.e., DMEM/F12 supplemented with 10% FBS, 2 mM L-glutamine and 100 μg ml−1 streptomycin. The cell culture was grown in tissue-culture treated Corning® flasks (Sigma-Aldrich, St. Louis, MO) in humidified atmosphere (5% CO2) at 37° C (Heraeus Hera Cell 150C incubator). For the cellular treatments, the day before the experiment cells were seeded at a density of 2·105 cells/mL in full medium on Corning® tissue-culture treated culture dishes (Sigma-Aldrich, St. Louis, MO). Immediately before use, stocks of phendione and cuproindione compounds were diluted (100x) in ultrapure H2O and then added to the cells in the culture medium at the desired final concentration (always less than 0.01% v/v of DMSO). For the cell experiments in copper-deprived medium, cells were pre-incubated with 50 μM of extracellular copper chelator BCS for three hrs and, maintaining the same medium, further incubated with the compounds.
6
Cell Culture and Transfection Protocols
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HeLa cells were grown in Dulbecco's modified Eagle Medium (Lonza) supplemented with 10% FBS, l -glutamine (2 mM), penicillin (100 U mL−1), streptomycin (100 mg mL−1). OCI-AML2 and OCI-AML3 [29] (link) cell lines were grown in MEM Alpha + GlutaMAX™-I medium (Gibco) supplemented with 20% FBS, glutamine and antibiotics. Transient transfections were performed using Lipofectamine™ 2000 (Invitrogen). Sf9 (Spodoptera frugiperda) insect cells were cultured at 27 °C in Sf 900 II SMF medium (Gibco) and transfected with pFastBacDual plasmids (Invitrogen) expressing either wild type NPM1 or NPMc+ using Insectogene T030-1.0 (Biontex). Baculoviral supernatant was collected after 96 h and used for two cycles of infection.
7
Cell Culture and Transfection Protocol
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U2OS and Flp-in 293 cells were cultured in Dulbecco’s modified Eagle medium (catalog no. 12-604F, Lonza) with 10% (volume [vol]/vol) fetal bovine serum at 37 °C with 5% CO2. The U2OS–TRE cells used for the DART system have been described in previous articles (28 (link)). For plasmid and siRNA transfection, Lipofectamine 2000 and Lipofectamine RNAiMax (Invitrogen) were used following the manufacturer’s standard protocol, respectively. The siRNA for TRDMT1 was purchased from Invitrogen (siRNA ID s4219; catalog no. 4392420). The siRNA for FMRP was purchased from Dharmacon (catalog no. L-019631-00-0005); the sequences of siRNA targeting FMRP 3′ UTR are 5′- GUACUGAGCAGUGAUAUUCdTdT-3′ and 5′- GAAUAUCACUGCUCAGUACdTdT-3′. Other siRNAs include siTET1 (catalog no. AM16708, assay ID:147892, Thermo), siBRCA1 (catalog no. L-003461-00, Dharmacon), and siBRCA2 (catalog no. GS675, Qiagen). Antibodies used in this study are summarized in SI Appendix , Table 1. Other methods used in the study are described in detail in SI Appendix , Materials and Methods.
8
RAW 264.7 Cell Culture Protocol
9
Cultivation of Colon Cancer Cell Lines
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Caco-2, T84, DLD-1, SW480, HT-29, and LS174T wildtype, colon adenocarcinoma cell line; HEK293, human embryonic kidney cell line; HeLa, cervical adenocarcinoma cell line; LS174T derived CDX2 knockout (LS174T CDX2 knockout) cell line [45 (link)] were grown in T175 culture flasks in Dulbecco’s Modified Eagle Medium (Lonza, Basel, Switzerland) or RPMI-1640 for DLD-1 (Thermo Fisher Scientific, Waltham, MA, USA), added 10% Fetal Bovine Serum gold (PAA) and 100 U/mL Penicillin-Streptomycin. Cells were incubated at 37 °C in 5% CO2 and passaged every 3–4 days when ~80% confluent. Passaging was done by removing media, rinsing three times with 85 mM sodium citrate, adding 1mL 0.05% trypsin EDTA (Invitrogen, Carlsbad, CA, USA) and incubating 5 min at 37 °C in 5% CO2.
10
Evaluating Clindamycin and Propolis Wound Healing
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Clindamycin hydrochloride was kindly gifted by European Egyptian pharmaceutical industry company, Alexandria, Egypt, and lyophilized red propolis extract was purchased from VACSERA, Giza, EGYPT, tea tree oil was purchased from Sigma – Aldrich, Mumbai, India. All other reagents and solvents were of HPLC analytical grade obtained from Fisher Scientific Company, USA.
Dulbecco's Modified Eagle Medium, penicillin–streptomycin (100×), fetal bovine serum, and phosphate-buffered saline were purchased from Lonza Group Ltd., Basel, Switzerland. Sulforhodamine-B and trisaminomethane base were purchased from Sigma-Aldrich, Louis, MO, USA. Trichloroacetic acid was purchased from Merck, Darmstadt, Germany.
Sprague–Dawley rats without skin damage or diseases were obtained from Misr university for science and technology animal center (Giza, Egypt). The ethical committee approved all animal and cell lines studies of Misr university for science and technology (Approval No: PH 7).
Dulbecco's Modified Eagle Medium, penicillin–streptomycin (100×), fetal bovine serum, and phosphate-buffered saline were purchased from Lonza Group Ltd., Basel, Switzerland. Sulforhodamine-B and trisaminomethane base were purchased from Sigma-Aldrich, Louis, MO, USA. Trichloroacetic acid was purchased from Merck, Darmstadt, Germany.
Sprague–Dawley rats without skin damage or diseases were obtained from Misr university for science and technology animal center (Giza, Egypt). The ethical committee approved all animal and cell lines studies of Misr university for science and technology (Approval No: PH 7).
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