The MG-63 cell line was used in this study because it can retain the differentiated phenotype over consecutive subcultures and shows a faster growth than the primary bone-forming lines, which makes it a good in vitro model. MG-63 cells (Sigma-Aldrich, Darmstadt, Germany) were cultivated in T75 culture flasks with Dulbecco’s Modified Eagle Medium (Biowest, Nuaillé, France) enriched with a 1% antibiotic (glutamine-penicillin-streptomycin [Biowest, Nuaillé, France]) and 10% foetal bovine serum (Biowest). After performing two cell subcultures and achieving an 80% confluence, the cells were seeded on the disc surfaces in a 24-microwells plate at a previously calculated concentration of 2 × 104 cells. Three discs were employed for each type of surface and the experiments were triplicated. Microwells without discs were used as negative controls. After 4 h of incubation, the cell culture was controlled by an Olympus CKX41SF2 phase contrast microscope (Olympus, Shinjuku-ku, Tokyo, Japan). Then, the microwells were completely filled with Dulbecco’s Modified Eagle Medium and incubated for a period of 24 h.
Dulbecco s modified eagle medium
Manufactured by Biowest
Sourced in France
Dulbecco's Modified Eagle Medium (DMEM) is a cell culture medium that provides essential nutrients and growth factors to support the in vitro cultivation of a variety of cell types. It is a widely used, standard formulation designed to maintain optimal pH and osmolarity for cell growth and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Biowest (2 mentions)
Glutamine penicillin streptomycin, by Biowest (1 mentions)
Mg 63 cells, by Merck Group (1 mentions)
Foetal bovine serum (fbs), by Biowest (1 mentions)
Hek293t, by Biowest (1 mentions)
Elisa, by BioLegend (1 mentions)
Hek293t, by Leibniz Institute DSMZ (1 mentions)
Skbr3, by American Type Culture Collection (1 mentions)
Hygromycin b, by Thermo Fisher Scientific (1 mentions)
Hepes, by Avantor (1 mentions)
Zeocin, by Thermo Fisher Scientific (1 mentions)
Penicillin g, by Merck Group (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Merck Group (1 mentions)
Blasticidin, by InvivoGen (1 mentions)
Farnesol, by Merck Group (1 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Biowest (1 mentions)
Glutamine, by Lonza (1 mentions)
Fetal bovine serum (fbs), by Biochrome (1 mentions)
7 protocols using dulbecco s modified eagle medium
1
MG-63 Cell Culture on Discs
2
Generating Engineered Cell Lines for Preclinical Studies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Green fluorescent protein and firefly luciferase-positive AsPC-1 cells (AsPC-1 WT) were generated in-house as described previously by Schaefer et al.28 (link) Luciferase expressing Raji cells were generated as described previously by Seitz et al.27 (link) AsPC-1 WT secreting latent TGF-β (AsPC-1-TGF) were generated via transduction of AsPC-1 WT with lentiviral vectors (LVV) encoding human TGF-β (UniProtKB-A0A024R0P8-1). The plasmid encoding human TGF-β was kindly provided by Stefan Edelburg (Miltenyi Biotec, Bergisch-Gladbach, Germany). Secretion of TGF-β was confirmed by ELISA (BioLegend, catalog no.: 432907). Human embryonic kidney-293 T cells (HEK293T, DSMZ no: Acc635) were obtained from DSMZ.
AsPC-1 WT and AsPC-1-TGF were cultured in RPMI 1640 (Biowest catalog no.: L0501) supplemented with 2 mM glutamine (Lonza, catalog no.: BE17-605E) and 10% fetal bovine serum (EXIMUS Maximus FBS, Catus Biotech, catalog no.: BS-2020-500). HEK293T cells were cultured in Dulbecco’s modified Eagle medium (Biowest, catalog no.: L0104) supplemented with 10% FBS. Cell lines and primary human cells were cultured at 37°C and 5% CO2.
AsPC-1 WT and AsPC-1-TGF were cultured in RPMI 1640 (Biowest catalog no.: L0501) supplemented with 2 mM glutamine (Lonza, catalog no.: BE17-605E) and 10% fetal bovine serum (EXIMUS Maximus FBS, Catus Biotech, catalog no.: BS-2020-500). HEK293T cells were cultured in Dulbecco’s modified Eagle medium (Biowest, catalog no.: L0104) supplemented with 10% FBS. Cell lines and primary human cells were cultured at 37°C and 5% CO2.
3
Culturing HER2-Positive Breast Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human Her2-overexpressing breast cancer cells SKBR3 (ATCC, Manassas, VA) were used in the study. Cells were maintained in Dulbecco’s Modified Eagle Medium (Biowest, Nuaillé, France) supplemented with 10% fetal bovine serum (Biowest, Nuaillé, France) and 80 µg/mL gentamycin (KRKA, Novo Mesto, Slovenia). Cells were grown at 37 °C in a humidified atmosphere containing 5% CO2.
4
Flp-In T-REx 293 Cell Culture Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Flp-In T-REx 293 human cell line (Thermofisher Scientific, Waltham, MA, USA), based on HEK293 cells, was cultured at 37 °C in 5% CO2 in an air‐ventilated humidified incubator according to manufacturer’s protocol, in high glucose Dulbecco’s Modified Eagle Medium (Biowest, Nuaillé, France) supplemented with 10% FBS, 1% Glutamax (Gibco), 10 mM HEPES (VWR LifeScience, Radnor, PA, USA), 100 U/mL penicillin G (Sigma Chemical Co., St. Louis, MO, USA), 80 U/mL streptomycin (Sigma), blasticidin (15 µg/mL) (InvivoGen, San Diego, CA, USA). The host cell line was maintained with zeocin (100 µg/mL) (InvitroGen, Carlsbad, CA, USA) and the transfected cell lines with hygromycin B (100 µg/mL) (InvitroGen). The experimental medium, the HEPES-buffered medium, (HBM: 137 mM NaCl, 5 mM KCl, 1.2 mM MgCl2, 0.44 mM KH2PO4, 4.2 mM NaHCO3, 1 mM CaCl2, 10 mM glucose, 20 mM HEPES, adjusted to pH 7.4 with NaOH), was used in all laboratory experiments. For the calcium elevation assay, HBM was supplemented with 1 mM probenecid [4-(dipropylsulfamoyl)benzoic acid] (Sigma) and 0.2% lipid-free bovine serum album.
5
Farnesol Modulates PMP22 in RT4 Schwann Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rat Schwann cells, RT4 (RT4-D6P2T, CRL-2768, ATCC; Manassas, VA, USA), were cultured in 10% fetal bovine serum, 1% penicillin-streptomycin solution (5000 units/mL penicillin, 5 mg/mL streptomycin), and low-glucose Dulbecco’s modified eagle medium (Biowest, Nuaille, France). RT4 cells (1 × 104) grown for 24 h in 12-well plates, transfected with either control and PMP22-containing vectors, pCMV-myc and pCMV-myc-PMP22 [34 (link)], respectively, using Lipofectamine 3000 reagent (Invitrogen, Paisley, UK) in accordance with the manufacturer’s protocol. Three hour after transfection, farnesol (Sigma, St. Louis, MO, USA) were treated onto the cells. To determine the effective concentration of farnesol, we have tested from 10 nM to 10 μM of farnesol, and 0.1 μM of farnesol was used in the data. The effect on the cell viability/proliferation was determined by direct counting under a microscope after 24, 48, and 72 h after farnesol treatment. To reduce the transfection variation, we performed at least three independent experiments.
6
Establishing Lymphoma and Melanoma Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For in vitro experiments, the Burkitt lymphoma cell line Raji as well as the melanoma cell line 526-Mel were transduced with lentiviral vectors encoding a GFP-firefly luciferase cassette. GFP+ 526-Mel cells were additionally modified to stably express CD20. For in vivo studies, a mouse-adapted RajiffLuc cell line was used [37 (link)]. All tumor cell lines were cultured in RPMI 1640 (Biowest, Nuaillé, France) supplemented with 2 mM glutamine (Lonza, Basel, Switzerland) and 10% fetal bovine serum (FBS, Biochrome, Berlin, Germany). The human embryonic kidney cell line 293 T was cultured in Dulbecco’s modified Eagle medium (Biowest, Nuaillé, France) supplemented with 10% FBS.
7
Evaluation of 5-FU and Fulvestrant
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following chemicals were used in this study: Dulbecco’s Modified Eagle Medium, and heat-inactivated fetal bovine serum (FBS) were purchased from Biowest (France), 5´-Fluorouracil, Fulvestrant, sodium dihydrogen phosphate, disodium hydrogen phosphate and gold (III) chloride trihydrate (HAuCl4·3H2O) were purchased from Sigma-Aldrich, USA. All reagents were of analytical grade.
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!