MIA PaCa-2, human PC cells, were cultured in DMEM (Lonza) containing L-Glutamine 2 mM, 10% heat-inactivated fetal bovine serum (FBS; Lonza) and 2,5% heat inactivated horse serum (HS; Lonza). PANC-1, human pancreatic epithelioid carcinoma cells, were kept in DMEM containing L-Glutamine 2 mM and 10% heat-inactivated fetal bovine serum (FBS; Lonza). BxPC-3, human pancreatic adenocarcinoma cells, were cultured in RPMI 1640 (Lonza) containing 10% heat-inactivated fetal bovine serum (FBS; Lonza). CAPAN-2, human PDAC cells, were kept in McCoy’s 5a Medium Modified (Lonza) with 10% heat-inactivated fetal bovine serum (FBS; Lonza). All the media were supplemented with antibiotics (10000 U/ml penicillin and 10 mg/ml streptomycin; Lonza). Cell lines were purchased from ATCC (Rockville, USA) and were grown at 37°C in 5% CO2 -95% air humidified atmosphere.
Mccoy s 5a medium modified
Manufactured by Lonza
Sourced in United Kingdom
McCoy's 5A Medium Modified is a cell culture medium formulated for the growth and maintenance of a variety of cell lines. It provides the necessary nutrients, growth factors, and other components to support cell proliferation and viability in in vitro cell culture applications.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Lonza (3 mentions)
Dulbecco modified eagle medium (dmem), by Lonza (2 mentions)
Rpmi 1640, by Lonza (2 mentions)
Streptomycin, by Lonza (2 mentions)
Fetal bovine serum (fbs), by Euroclone (2 mentions)
Iscove modified dulbecco media (imdm), by Lonza (2 mentions)
A2780, by European Collection of Cell Cultures (2 mentions)
Streptomycin p s, by Lonza (2 mentions)
Mia paca 2, by American Type Culture Collection (2 mentions)
Aspc 1, by American Type Culture Collection (2 mentions)
Tissue culture flasks, by Sarstedt (2 mentions)
Skov3, by European Collection of Cell Cultures (2 mentions)
Roswell park memorial institute (rpmi), by Lonza (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
L glutamine, by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Lonza (1 mentions)
Penicillin streptomycin, by Lonza (1 mentions)
Htb 4, by American Type Culture Collection (1 mentions)
Verapamil, by Merck Group (1 mentions)
Roswell park memorial institute (rpmi), by Corning (1 mentions)
10 protocols using mccoy s 5a medium modified
1
Culturing Pancreatic Cancer Cell Lines
2
Inflammatory Response in Bladder Epithelial Cells
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T24 human bladder epithelial cells (HTB-4; American Type Culture Collection, USA) were maintained in McCoy’s 5A Medium Modified (Lonza, Ltd., Basel, Switzerland) supplemented with 10% fetal bovine serum (FBS, EuroClone S.p.A, Pero, Italy) and 1% penicillin/streptomycin (Lonza Ltd., Basel, Switzerland).
T24 cells were seeded in 6-well plates and cultured for 24 h before treatments. The inflammation was induced using TNFα treatment (10 ng/mL, Shenandoah Biotechnology Inc. Warminster PA, USA) for 6 h and 24 h. TNFα concentration was determined by concentration dependence experiments followed by the analysis of pro-inflammatory cytokine mRNA expression (Supplementary Fig.2 .) The anti-inflammatory effects of EOs and their main components were determined in three different experiments: TNFα pre-treatment for 6 h followed by 24 h of EO/standard treatment; TNFα pre-treatment for 24 h followed by 6 h of EO/standard treatment; and TNFα pre-treatment for 24 h followed by 24 h of EO/standard treatment. The EOs and standards were used at 500-fold dilution of stock solutions to determine their effect on cytokine production. In all experiments, cells treated with DMSO were used as controls. All experiments were performed at least 3 times and carried out at 37 °C in a humidified atmosphere containing 5% CO2.
T24 cells were seeded in 6-well plates and cultured for 24 h before treatments. The inflammation was induced using TNFα treatment (10 ng/mL, Shenandoah Biotechnology Inc. Warminster PA, USA) for 6 h and 24 h. TNFα concentration was determined by concentration dependence experiments followed by the analysis of pro-inflammatory cytokine mRNA expression (Supplementary Fig.
3
Cytotoxicity Evaluation of Doxorubicin and Verapamil
4
Cell Culture of Cancer Cell Lines
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The cancer cell lines AsPC-1 and MIA PaCa-2 were purchased from the American Type Culture Collection (UK). The cancer cell lines A2780 and SK-OV-3 were purchased from the European Collection of Cell Cultures (UK). All cells were grown using distributors' instructions. All cells were cultured in either IMDM, McCoy's 5a Medium Modified or RPMI (Lonza, UK) substituted with 10% FBS (15% for McCoy's 5a Medium Modified) (Bio-Sera, UK) and (v/v); 100 units/mL penicillin, 100 µg/mL streptomycin (P/S) (Lonza, UK). All serum was filtered using a 0.2 µM syringe filter prior to addition to media. When not in use all media was stored between 4-6 °C. All cells were incubated at 37 °C in a 5% CO2 atmosphere. Cells were cultured in tissue culture flasks (Sarstedt, UK) and removed via scraping when cells were 70-90% confluent.
5
Cell Line Cultivation and Maintenance
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The cell lines AsPC-1, Caco-2, Colo320, HCT116, JJN3, Lovo, MCF7, MDA-MB-231, MIA PaCa-2, MM.1S and U266B1 were purchased from the American Type Culture Collection (UK). The cell lines A2780, SK-OV-3, T47D and U937 were purchased from European Collection of Cell Cultures (UK). All cell lines were cultured in accordance with distributor's recommendations. Each cell line was cultured using either DMEM, RPMI, IMDM or McCoy's 5a Medium Modified (Lonza, UK) with FBS (Bio-Sera, UK) and (v/v); 100 units/ml penicillin, 100 µg/ml streptomycin (P/S) (Lonza, UK). All serum, P/S and buffers added to media were filtered through a 0.2µm filter before addition. Between use all media was stored at 2-8°C. All cell lines were incubated at 37°C in a 5% CO2 atmosphere except for MDA-MB-231 which was incubated at 37°C in a 0% CO2 atmosphere. All cell lines were cultured in tissue culture flasks (Sarstedt, UK) and removed when cells were either adherent and in the logarithmic growth phase or removed when at a high enough growth density for suspension cell lines.
6
Cell Culture Conditions for BJ, U2OS, and U2OS-R1 Cells
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BJ cells (CRL-2522) were grown in Eagle’s
Minimum Essential Medium from Sigma-Aldrich. U2OS (HTB-96) and U2OS
stably transfected with FGFR1 (U2OS-R1) were grown in McCoy’s
5A Modified Medium from Lonza (Switzerland). All media were supplemented
with 10% fetal bovine serum from Thermo Fisher Scientific, and 1%
penicillin/streptomycin mix was from BioWest (France). Additionally,
the U2OS-R1 cell medium contained 50 μg/mL gentamicin sulfate
from Thermo Fisher Scientific. All cell lines were cultured in a humidified
incubator at 37 °C in 5% CO2 atmosphere. The BJ and
U2OS cell lines were obtained from American Type Culture Collection
(Manassas, VA). The U2OS cells stably expressing FGFR1 (U2OS-R1) were
a kind gift from Dr. Ellen M. Haugsten from The Norwegian Radium Hospital.44 (link)
Minimum Essential Medium from Sigma-Aldrich. U2OS (HTB-96) and U2OS
stably transfected with FGFR1 (U2OS-R1) were grown in McCoy’s
5A Modified Medium from Lonza (Switzerland). All media were supplemented
with 10% fetal bovine serum from Thermo Fisher Scientific, and 1%
penicillin/streptomycin mix was from BioWest (France). Additionally,
the U2OS-R1 cell medium contained 50 μg/mL gentamicin sulfate
from Thermo Fisher Scientific. All cell lines were cultured in a humidified
incubator at 37 °C in 5% CO2 atmosphere. The BJ and
U2OS cell lines were obtained from American Type Culture Collection
(Manassas, VA). The U2OS cells stably expressing FGFR1 (U2OS-R1) were
a kind gift from Dr. Ellen M. Haugsten from The Norwegian Radium Hospital.44 (link)
7
Synergistic Effects of I3C and Aspirin
8
Culturing Human Adenocarcinoma and Monocyte Cells
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Human colon adenocarcinoma cell line HT-29 (Cat. No. 91072201) and human monocyte-like THP-1 cells (Cat. No. 88081201) were purchased from the European Collection of Authenticated Cell Cultures (ECACC) and cultured in McCoy’s 5A Modified Medium (Bio Whittaker Lonza, Verviers, Belgium) and RPMI-1640 (Bio Whittaker Lonza, Verviers, Belgium) respectively; they were supplemented with 10% fetal bovine serum (FBS, Euroclone, Milan, Italy) and 100 IU/mL penicillin + 100 μg/mL streptomycin (Lonza, Basel, Switzerland) (complete culture medium). Cell lines were incubated at 37 °C in an atmosphere supplemented with 5% CO2, in 75 cm2 flasks. The adherent cell line, HT-29, was cultured until ~85% confluence; it was then washed with phosphate-buffered saline (PBS, Merck, Darmstadt, Germany), and detached using 0.05% trypsin-EDTA solution (Thermo Fischer Scientific, Waltham, MA, USA). The cells were then suspended in a complete culture medium, washed by centrifugation at 200× g for 10 min, and then re-suspended in a fresh complete culture medium. The non-adherent cell lines (THP-1) were simply collected and centrifuged in the previously mentioned conditions.
9
Cell Culture Maintenance and METTL11A KO
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Human embryonic kidney (HEK293T) cells were maintained in Dulbecco’s Modified Eagle Medium (Corning Incorporated) supplemented in 10% fetal bovine serum (R&D Systems, Inc) and 1% penicillin-streptomycin (P/S, Corning Incorporated). HCT116 human colorectal carcinoma and CRISPR/Cas9-mediated METTL11A KO HCT116 cell lines were cultured in McCoy’s 5A Modified Medium (Lonza) supplemented with 10% fetal bovine serum and 1% P/S. METTL11A KO cells were generated previously (10 (link)). Maintenance of METTL11A loss was verified by Western blot (Fig. 6 B). HEK293T and HCT116 cell lines were a generous gift from Dr Ian Macara.
10
Breast Cancer Cell Line Culture
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Cell lines were obtained from DSMZ Cell Culture Collection or ATCC. CAL-51 (ACC 302), MCF-7 (ACC 115) and ZR-75-1 (CRL-1500) cell lines were cultured in high glucose Dulbecco’s Modified Eagle Medium (DMEM) containing L-glutamine (Sigma Aldrich, UK). The MDA-MB-468 (HTB-132) and T-47D (HTB-133) cell lines were cultured in Roswell Park Memorial Institute (RPMI)-1640 medium containing L-glutamine (Sigma Aldrich, UK). The SK-BR-3 (HTB-30) cell line was cultured in high glucose McCoy’s 5A (modified) medium containing L-glutamine (Lonza, CH). All media was supplemented with 10% foetal calf serum and 1x non-essential amino acids (Sigma Aldrich, UK).
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