Muller (ENW001, Kerafast, Boston, MA) and R28 (EUR201, Kerafast) cells were cultured in 5% CO2 humidified incubator at 37°C. The culture media for Muller cells consisted of DMEM (10–017-CV, Corning, Corning, NY) supplemented with final concentrations of 1% Pen-Strep (17-6020E, Lonza, Basel, Switzerland), 10% fetal bovine serum (16140–071, Gibco, Waltham, MA), and additional 1% L-glutamine (3772, Carl Roth, Karlsruhe, Germany). The culture media for the retinal cell line R28 consisted of the following: DMEM (10-013-CV, Corning) supplemented with 1× MEM nonessential amino acids (11140-050, Gibco), 1% Pen-Strep, 10% bovine calf serum (30–2030, ATCC, Manassas, VA), sodium bicarbonate (15 mL of 7.5 w/v %, 7412-12, Mallinckrodt Chemicals, St. Louis, MO), and L-glutamine (5 mL of 200 mM stock, 3772, Carl Roth). We used 1× PBS to dilute Trypsin-EDTA (25-053-CI, Corning) 1:3 for Muller cells. CMF-EDTA was used as the trypsin dilutant for R28 cells. The recipe for CMF-EDTA was followed as outlined by the R28 care document from Kerafast.
Dmem 10 017 cv
Manufactured by Corning
Sourced in United States
DMEM (10-017-CV) is a basal medium designed for the cultivation of a variety of cell types. It provides the essential nutrients and supplements required for cell growth and maintenance in in vitro cell culture applications.
Automatically generated - may contain errors
Lab products found in correlation
Heparin, by Merck Group (2 mentions)
Penicillin streptomycin pen strep, by Merck Group (2 mentions)
Hepes buffer, by Merck Group (2 mentions)
Dulbecco s phosphate buffered saline, by Thermo Fisher Scientific (2 mentions)
Mem non essential amino acid solution, by Merck Group (2 mentions)
2 mercaptoethanol, by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Atlas Biologicals (2 mentions)
Cellometer k2 cell counter, by Revvity (2 mentions)
Bovine calf serum (bcs), by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dmem 10 013 cv, by Corning (1 mentions)
Pen strep, by American Type Culture Collection (1 mentions)
L glutamine, by Carl Roth (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Corning (1 mentions)
Pen strep, by Lonza (1 mentions)
Equafetal, by Atlas Biologicals (1 mentions)
4 protocols using dmem 10 017 cv
1
Culturing Muller and R28 Retinal Cell Lines
2
MDBK Cell Line Maintenance Protocol
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The Madin Darby bovine kidney (MDBK) cell line was maintained in Dulbecco’s modified Eagles medium (DMEM # 10-017-CV, Corning®, Corning, NY, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS; EquaFETAL, Atlas Biologicals, Fort Collins, CO, USA) and 1× antibiotic/antimycotic solution (cat. # 30-004-CI; Corning®).
3
Isolation of Murine Lung Single-Cell Suspension
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As described previously, mice were euthanized and lungs perfused with 10 ml 1× Dulbecco's phosphate-buffered saline without calcium or magnesium (Gibco) (PBS) containing 50 U/ml heparin (Sigma) and placed into 2 ml complete Dulbecco’s modified Eagle’s medium (c-DMEM), DMEM (10-017-CV, Corning), 500 ml supplemented with filter-sterilized 5 ml HEPES buffer (1 M; Sigma), 10 ml MEM nonessential amino acid solution (100×; Sigma), 5 ml penicillin-streptomycin (pen/strep) (100×; Sigma), 660 μl 2-mercaptoethanol (50 mM; Sigma), and 45 ml heat-inactivated fetal bovine serum (FBS) (Atlas Biologicals). A single-cell suspension was obtained using enzymatic digestion (49 (link)). Residual erythrocytes were lysed using Gey’s solution (8 mM NH4Cl, 5 mM KHCO3 in water), passed through a 40-μm strainer, and suspended in c-DMEM. The total number of viable cells was determined with acridine orange and propidium iodide (AO/PI) staining and counted on a Cellometer K2 cell counter (Nexcelom Bioscience).
4
Isolation of Murine Lung Single-Cell Suspension
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As described previously, mice were euthanized and lungs perfused with 10 ml 1× Dulbecco's phosphate-buffered saline without calcium or magnesium (Gibco) (PBS) containing 50 U/ml heparin (Sigma) and placed into 2 ml complete Dulbecco’s modified Eagle’s medium (c-DMEM), DMEM (10-017-CV, Corning), 500 ml supplemented with filter-sterilized 5 ml HEPES buffer (1 M; Sigma), 10 ml MEM nonessential amino acid solution (100×; Sigma), 5 ml penicillin-streptomycin (pen/strep) (100×; Sigma), 660 μl 2-mercaptoethanol (50 mM; Sigma), and 45 ml heat-inactivated fetal bovine serum (FBS) (Atlas Biologicals). A single-cell suspension was obtained using enzymatic digestion (49 (link)). Residual erythrocytes were lysed using Gey’s solution (8 mM NH4Cl, 5 mM KHCO3 in water), passed through a 40-μm strainer, and suspended in c-DMEM. The total number of viable cells was determined with acridine orange and propidium iodide (AO/PI) staining and counted on a Cellometer K2 cell counter (Nexcelom Bioscience).
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