Mouse melanoma cell lines SW1 and PVMM were obtained as generous gifts from Dr. Ze’ev Ronai of Sanford Burnham Prebys Medical Discovery Institute and Dr. Serge Y. Fuchs of University of Pennsylvania, respectively. Cells were maintained as monolayers in Dulbecco’s Modified Eagle’s Medium (VWR International), supplemented with 10% v/v fetal bovine serum (FBS, Gibco by Life Technologies), 100 IU of Penicillin G and 100 μg/ml Streptomycin (Corning). The cell lines were tested for mycoplasma contamination with MycoAlert Plus Mycoplasma detection kit (Lonza).
Dulbecco s modified eagle s medium
Manufactured by Avantor
Dulbecco's Modified Eagle's Medium (DMEM) is a commonly used cell culture medium that supports the growth and maintenance of various cell types. It provides a balanced salt solution, amino acids, vitamins, and other nutrients essential for cell proliferation and survival.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Corning (2 mentions)
Penicillin g, by Corning (2 mentions)
Mycoalert plus mycoplasma detection kit, by Lonza (2 mentions)
Optima grade methanol, by Thermo Fisher Scientific (2 mentions)
Formic acid, by Thermo Fisher Scientific (2 mentions)
Acetonitrile, by Thermo Fisher Scientific (2 mentions)
Mgcl2, by Avantor (2 mentions)
7 oh cbd, by Cayman Chemical (2 mentions)
Geneticin g418, by Avantor (2 mentions)
Diclofenac, by Merck Group (2 mentions)
Nicotine, by Merck Group (2 mentions)
Trifluoperazine, by Merck Group (2 mentions)
Dulbecco s phosphate buffered saline, by Avantor (2 mentions)
Ultra low binding microcentrifuge tubes, by Avantor (2 mentions)
Hela cell, by American Type Culture Collection (1 mentions)
Chenodeoxycholic acid, by Merck Group (1 mentions)
Serotonin, by Merck Group (1 mentions)
Propofol, by Merck Group (1 mentions)
Acetaminophen, by Merck Group (1 mentions)
6 protocols using dulbecco s modified eagle s medium
1
Culturing Mouse Melanoma Cell Lines
2
Culturing Mouse Melanoma Cell Lines
3
Automated Cell Culture and Dispensing
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Cell Culture and Dispensing.
HeLa cells were obtained from the American Type Culture Collection (Manassas, VA) and cultured at 37 °C with 5% CO 2 . The cells were maintained using Dulbecco's Modified Eagle's Medium (VWR, Radnor, PA) with 10% fetal bovine serum and 1% penicillin/streptomycin. The cells were split and harvested upon reaching ~90% confluency as described previously. 35 Before use, cultured HeLa cells were filtered using a 40 μm FACS Tube Cap filter (Stellar Scientific, Baltimore, MD) and washed three times in phosphate-buffered saline (PBS) (Corning, Corning, NY) with centrifugation at 200 × g for 5 min prior to each wash. The cells were then resuspended in PBS to a final concentration of approximately 3 × 10 5 cells/mL, loaded into a piezoelectric dispensing capillary in the cellenONE X1 (Cellenion, Lyon, France), and gated to represent the majority population.
HeLa cells were obtained from the American Type Culture Collection (Manassas, VA) and cultured at 37 °C with 5% CO 2 . The cells were maintained using Dulbecco's Modified Eagle's Medium (VWR, Radnor, PA) with 10% fetal bovine serum and 1% penicillin/streptomycin. The cells were split and harvested upon reaching ~90% confluency as described previously. 35 Before use, cultured HeLa cells were filtered using a 40 μm FACS Tube Cap filter (Stellar Scientific, Baltimore, MD) and washed three times in phosphate-buffered saline (PBS) (Corning, Corning, NY) with centrifugation at 200 × g for 5 min prior to each wash. The cells were then resuspended in PBS to a final concentration of approximately 3 × 10 5 cells/mL, loaded into a piezoelectric dispensing capillary in the cellenONE X1 (Cellenion, Lyon, France), and gated to represent the majority population.
4
In Vitro Metabolism of Cannabinoids
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THC, 11-OH-THC, THC-COOH, THC-COO-Gluc, CBD, 7-OH-CBD, and CBN were purchased from Cayman Chemicals (Ann Arbor, MI) or Sigma-Aldrich (St. Louis, MO). Pooled human liver microsomes (HLMs) [n = 50, mixed gender (21 female and 29 male), race (42 Caucasian, 4 Hispanic, 2 African American, and 2 Asian), and age (5–77 years)] and pooled human kidney microsomes (HKMs) [n = 8, mixed gender (50% each), race (3 African American, 3 Caucasian, and 2 Hispanic), and age (42–70 years)] were obtained from Sekisui Xenotech, LLC (Lenexa, KS). β-estradiol, chenodeoxycholic acid, trifluoperazine, serotonin, propofol, codeine, zidovudine (AZT), nicotine, oxazepam, dihydroexemestane, ketoconazole, diclofenac, acetaminophen, and furosemide were all purchased from Sigma-Aldrich. Optima grade methanol, acetonitrile, and formic acid were obtained from Fisher Scientific (Waltham, MA). Ultra-low-binding microcentrifuge tubes, Dulbecco’s modified Eagle’s medium, Dulbecco’s phosphate-buffered saline, UDPGA, alamethacin, MgCl2, and geneticin (G418) were purchased from VWR (Radnor, PA). BCA protein assays were purchased from Pierce (Rockford, IL), premium grade FBS was purchased from Seradigm (Radnor, PA), and ChromatoPur bovine serum albumin (BSA) was purchased from MB Biomedicals (Santa Ana, CA).
5
Caco-2 Epithelial Monolayer: Toxin Exposure
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Caco-2 cells (ATCC HTB-37) were grown in Dulbecco’s Modified Eagle’s Medium (VWR, Vienna, Austria) supplemented with 10% FCS, 2 mM glutamine (VWR, Vienna, Austria), 50 mg/l each penicillin and streptomycin (VWR, Vienna, Austria) and 1% non-essential amino-acids (VWR, Vienna, Austria). Cells were seeded onto 24-well Transwell™ membrane inserts (Corning, USA) placed in 24-well companion plates (Corning, USA), 24-well plates (for resazurin assay) or 3 cm Ø dishes (for glucosylation assay) and grown until day 21 after confluence to form a monolayer of polarized epithelial cells. In all incubation solutions containing clostridial toxins, the FCS concentration was lowered to 0.1%.
6
Quantitative Analysis of CBD and 7-OH-CBD Metabolism
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CBD and 7-OH-CBD were purchased
from Cayman Chemicals (Ann Arbor, MI) or Sigma-Aldrich (St. Louis,
MO). Pooled human liver microsomes (HLM) [n = 50,
mixed-gender (21 female and 29 male), race (42 Caucasian, 4 Hispanic,
2 African American, and 2 Asian), and age (5–77 years old)]
were obtained from Sekisui Xenotech, LLC (Lenexa, Kansas). NADPH-regenerating
system (1.3 mM NADP, 3.3 mM glucose 6-phosphate, and 0.4 U/mL glucose
6-phosphate dehydrogenase) was obtained from Corning (Bedford, MA).
Nicotine, norNicotine, HPBA, NOX, Nic-Gluc, cotinine, COX, Cot-Gluc,
3HC, and 3HC glucuronide (3HC-Gluc) standards were purchased from
Sigma-Aldrich. Fluconazole, benzydamine hydrochloride, clopidogrel,
trifluoperazine, diclofenac, amitriptyline, and tranylcypromine were
also purchased from Sigma-Aldrich. Optima grade methanol, acetonitrile,
and formic acid were obtained from Fisher Scientific (Waltham, MA).
Ultra-low-binding microcentrifuge tubes, Dulbecco’s modified
Eagle’s medium, Dulbecco’s phosphate-buffered saline,
UDP glucuronic acid (UDPGA), alamethicin, MgCl2, and Geneticin
(G418) were purchased from VWR (Radnor, PA). BCA protein assays were
purchased from Pierce (Rockford, IL); premium-grade fetal bovine serum
(FBS) was purchased from Seradigm (Radnor, PA), and ChromatoPur bovine
serum albumin (BSA) was purchased from MB Biomedicals (Santa Ana,
CA).
from Cayman Chemicals (Ann Arbor, MI) or Sigma-Aldrich (St. Louis,
MO). Pooled human liver microsomes (HLM) [n = 50,
mixed-gender (21 female and 29 male), race (42 Caucasian, 4 Hispanic,
2 African American, and 2 Asian), and age (5–77 years old)]
were obtained from Sekisui Xenotech, LLC (Lenexa, Kansas). NADPH-regenerating
system (1.3 mM NADP, 3.3 mM glucose 6-phosphate, and 0.4 U/mL glucose
6-phosphate dehydrogenase) was obtained from Corning (Bedford, MA).
Nicotine, norNicotine, HPBA, NOX, Nic-Gluc, cotinine, COX, Cot-Gluc,
3HC, and 3HC glucuronide (3HC-Gluc) standards were purchased from
Sigma-Aldrich. Fluconazole, benzydamine hydrochloride, clopidogrel,
trifluoperazine, diclofenac, amitriptyline, and tranylcypromine were
also purchased from Sigma-Aldrich. Optima grade methanol, acetonitrile,
and formic acid were obtained from Fisher Scientific (Waltham, MA).
Ultra-low-binding microcentrifuge tubes, Dulbecco’s modified
Eagle’s medium, Dulbecco’s phosphate-buffered saline,
UDP glucuronic acid (UDPGA), alamethicin, MgCl2, and Geneticin
(G418) were purchased from VWR (Radnor, PA). BCA protein assays were
purchased from Pierce (Rockford, IL); premium-grade fetal bovine serum
(FBS) was purchased from Seradigm (Radnor, PA), and ChromatoPur bovine
serum albumin (BSA) was purchased from MB Biomedicals (Santa Ana,
CA).
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