Chinese hamster ovary (CHO) cells stably expressing M23-AQP4 (CHO-AQP4 cells) were used, as described (Crane et al., 2011 (link); Phuan et al., 2013 (link)). CHO cells were cultured at 37 °C in 5% CO2 95% air in F-12 Ham’s Nutrient Mixture medium supplemented with 10% fetal bovine serum, 200 μg/ml geneticin, 100 U/ml penicillin and 100 μg/ml streptomycin. Human natural killer cells (NK cells) expressing the high-affinity 176V variant of the Fcγ receptor IIIA were obtained from Fox Chase Cancer Center (Philadelphia, PA). NK cells were cultured in suspension in α-MEM (Sigma-Aldrich, St. Louis, MO) containing 0.1 mM 2-mercaptoethanol, 2 mM L-glutamine, 0.2 mM myoinositol, 10% FBS, 10% horse serum, 2.5 μM folic acid, non-essential amino acids, 1 mM Na pyruvate, 100 U/ml penicillin, 100 μg/ml streptomycin, and 100 IU/ml of human recombinant interleukin-2 (GenScript, Piscataway, NJ). Primary astrocyte cultures were generated from cerebral cortex of neonatal rats as described (Duan et al., 2018 (link)).
Penicillin
Manufactured by GenScript
Penicillin is a type of laboratory equipment used for the cultivation and storage of microorganisms, particularly bacteria. It serves as a container to maintain the appropriate environmental conditions, such as temperature and humidity, required for the growth and preservation of these microbial samples.
Automatically generated - may contain errors
Lab products found in correlation
Streptomycin, by GenScript (3 mentions)
M csf, by GenScript (2 mentions)
α mem, by Merck Group (1 mentions)
Dispase 1, by Merck Group (1 mentions)
Penicillin, by American Type Culture Collection (1 mentions)
Streptomycin, by American Type Culture Collection (1 mentions)
0.45 μm strainer, by Corning (1 mentions)
Penicillin, by Merck Group (1 mentions)
Streptomycin, by Merck Group (1 mentions)
Lipopolysaccharides (lps), by InvivoGen (1 mentions)
3 protocols using penicillin
1
Stably Expressing CHO Cells for NK Cell Interaction
2
Differentiation and Culture of Primary Mouse Cell Types
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BMDM were differentiated from bone marrow incubated in DMEM with 10% fetal calf serum (FCS), 2 mM L-glutamine, 10U/ml Penicillin, 10 μg/ml Streptomycin, and 10 ng/ml M-CSF (GenScript) for 7 days.
NIH 3T3 cells (mouse, male, ATCC CRL-1658) were maintained in DMEM supplemented with 10% Newborn Calf Serum (NBCS), 2 mM L-glutamine, 10U/ml Penicillin, and 10 μg/ml Streptomycin.
Primary corneal epithelial cells were collected from C57Bl/6 mice by incubating eyes overnight at 4 °C in a 1:1 mixture of DMEM:F12 supplemented with 4 mg/ml Dispase I (Sigma–Aldrich), 10U/ml Penicillin, 10 μg/ml Streptomycin, and 0.025 μg/ml Amphotericin B. Epithelial sheets were peeled from eyes and dissociated in TrypLE for 10 min at 37 °C with gentle agitation. Cells were passed through a 0.45 μm strainer (Corning), washed, and cultured on collagen and fibronectin-coated plates in KSFM supplemented with 5 ng/ml hEGF, 50 μg/ml Bovine Pituitary Extract, and 100 ng/ml Cholera Toxin.
All cells were maintained at 37 °C with 5% CO2. Adherent cells were detached for subculture and experiments using TrypLE. Unless otherwise noted, all cell culture materials were purchased from Gibco/Thermo Fisher.
NIH 3T3 cells (mouse, male, ATCC CRL-1658) were maintained in DMEM supplemented with 10% Newborn Calf Serum (NBCS), 2 mM L-glutamine, 10U/ml Penicillin, and 10 μg/ml Streptomycin.
Primary corneal epithelial cells were collected from C57Bl/6 mice by incubating eyes overnight at 4 °C in a 1:1 mixture of DMEM:F12 supplemented with 4 mg/ml Dispase I (Sigma–Aldrich), 10U/ml Penicillin, 10 μg/ml Streptomycin, and 0.025 μg/ml Amphotericin B. Epithelial sheets were peeled from eyes and dissociated in TrypLE for 10 min at 37 °C with gentle agitation. Cells were passed through a 0.45 μm strainer (Corning), washed, and cultured on collagen and fibronectin-coated plates in KSFM supplemented with 5 ng/ml hEGF, 50 μg/ml Bovine Pituitary Extract, and 100 ng/ml Cholera Toxin.
All cells were maintained at 37 °C with 5% CO2. Adherent cells were detached for subculture and experiments using TrypLE. Unless otherwise noted, all cell culture materials were purchased from Gibco/Thermo Fisher.
3
Differentiation and Activation of Bone Marrow-Derived Macrophages
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BMDM were differentiated from bone marrow incubated in DMEM with 10% fetal calf serum (FCS), 2mM L-glutamine, 10U/ml Penicillin, 10μg/ml Streptomycin, and 10ng/ml M-CSF (GenScript) for 7 days. BMDM were activated by priming overnight with 10nM IFNɣ (Peprotech), followed by 5 hours with 200ng/ml LPS (Invivogen).
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