MHCC‐97H, PLC/PRF/5, HEK293, and HepG2 cells were cultured in DMEM (HyClone, Logan, UT, USA) mixed with 1% penicillin/streptomycin (HyClone) and 10% fetal bovine serum (FBS, Gibco, NY, USA) at 37 °C with 5% CO2. SNU449 cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Gibco, Grand Island, NY, USA) mixed with 1% penicillin/streptomycin (MedChemExpress, Monmouth, NJ, USA) and 10% FBS at 37 °C with 5% CO2.
Penicillin streptomycin
Manufactured by MedChemExpress
Sourced in United States, China
Penicillin/streptomycin is a commonly used antibiotic solution that contains a combination of penicillin and streptomycin. It is a broad-spectrum antibiotic often used in cell culture applications to prevent bacterial contamination.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Fetal bovine serum (fbs), by MedChemExpress (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Chir99021, by Selleck Chemicals (1 mentions)
A83 01, by MedChemExpress (1 mentions)
Fibroblast growth factor basic bfgf, by Thermo Fisher Scientific (1 mentions)
Low glucose dmem, by Thermo Fisher Scientific (1 mentions)
Y 27632, by Selleck Chemicals (1 mentions)
Basic fibroblast growth factor (bfgf), by MedChemExpress (1 mentions)
Scc25, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Cyclopamine, by MedChemExpress (1 mentions)
Regorafenib, by Cayman Chemical (1 mentions)
Lenvatinib, by Cayman Chemical (1 mentions)
Protease inhibitor cocktail, by Selleck Chemicals (1 mentions)
Iodoacetamide iam, by Merck Group (1 mentions)
Sorafenib, by Cayman Chemical (1 mentions)
Bicinchoninic acid bca assay, by Solarbio (1 mentions)
7 protocols using penicillin streptomycin
1
Cell Culture Protocol for Various Cell Lines
2
Expansion and Characterization of Expanded ACY-Treated MSCs
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In an ACY (-) group, MSCs were cultured as previously described (Henrionnet et al., 2017 (link); Samal et al., 2021 (link)). In brief, the culture medium contained low glucose DMEM (Gibco) supplemented with FBS (10%, Hyclone), fibroblast growth factor-basic (bFGF, 1 ng/ml, PeproTech), and penicillin/streptomycin (1%, Gibco). In ACY (+) group, the basal medium for culturing MSCs consisted of low glucose DMEM, FBS, bFGF, penicillin/streptomycin, and ACY cocktails consisted of A-83–01 (MedChemExpress), CHIR99021 (Selleck), and Y-27632 (Selleck). The medium was changed 1 day after seeding and every 2 days thereafter. There were 2 μM A-83–01, 3 μM CHIR99021, and 2 μM Y-27632 chosen as the optimal concentration in the ACY (+) group. When cells were approximately 90% confluent, they would be passaged with a split ratio of 1:3.
MSCs at passage 2 (P2) were seeded, without (ACY-) or treated (ACY+) with A-83–01, CHIR99021, and Y-27632, and were continuously cultured to passage 10 (P10;Supplementary Figure S1 ). From passage 5 (P5) to P10, the MSCs were used in subsequent analysis.
MSCs at passage 2 (P2) were seeded, without (ACY-) or treated (ACY+) with A-83–01, CHIR99021, and Y-27632, and were continuously cultured to passage 10 (P10;
3
Cell Culture Protocols for Cancer Research
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The TU686 and SCC25 cell lines were purchased from The iCell Bioscience Inc,Shanghai (iCell-h300 and iCell-h361, Shanghai, China). The WSU-HN30 cell lines were purchased from Shanghai Zephyr Biotechnology Co. (ZYH60432,Shanghai, China).
The cell TU686 lines were cultured in RPMI 1640 (Gibco, New York, NY, United States) with 10% fetal bovine serum (FBS, Gibco), while SCC25 and WSU-HN30 cell lines were cultured in DMEM (Gibco, New York, NY, United States) supplemented with 10% FBS (Gibco) serum. These cell lines were cultured in a medium supplemented with penicillin–streptomycin (each at 100 Units/ml, MedChemExpress, Shanghai, China) at 37 °C in an incubator with humidifified atmosphere and 5% CO2.
The cell TU686 lines were cultured in RPMI 1640 (Gibco, New York, NY, United States) with 10% fetal bovine serum (FBS, Gibco), while SCC25 and WSU-HN30 cell lines were cultured in DMEM (Gibco, New York, NY, United States) supplemented with 10% FBS (Gibco) serum. These cell lines were cultured in a medium supplemented with penicillin–streptomycin (each at 100 Units/ml, MedChemExpress, Shanghai, China) at 37 °C in an incubator with humidifified atmosphere and 5% CO2.
4
Osteoblast Behavior on Titanium Surfaces
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The human MG63 osteoblasts were obtained from ATCC (Rockville, MD, USA). The human MG63 osteoblasts were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 1% antibiotic mixture (penicillin/streptomycin; HyClone, Logan, UT, USA) and 10% fetal bovine serum (FBS; Thermo Fisher Scientific) at 37°C under a 5% CO2 humidified atmosphere. The MG63 osteoblasts were seeded onto the four different surfaces of titanium disks in 12-well plates with a concentration of 5×104/well. The cyclopamine (MedChem Express, Princeton, NJ, USA) was dissolved at 5 mg/mL in dimethyl sulphoxide, and the solution was diluted to the final concentration with DMEM supplemented with 1% antibiotic mixture (penicillin/streptomycin) and 10% FBS. Treatment with control vehicle was used for control cells.
5
Comparative Efficacy of Sorafenib, Regorafenib, and Lenvatinib
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Sorafenib, regorafenib and lenvatinib were from Cayman Chemical Company (Michigan, United States); DMSO and bicinchoninic acid (BCA) assay were from Solarbio Science and Technology Company (Beijing, China); fetal bovine serum (FBS), Dulbecco’s modified essential media (DMEM) and trypsin-EDTA were from Gibco (California, United States); cell counting kit-8 (CCK-8) was from Dojindo Corporation (Shanghai, China); penicillin-streptomycin was from MedChem Express (New Jersey, United States); dithiothreitol (DTT) and Tris were from BBI Life Sciences (Shanghai, China); rLys-C and trypsin were from Promega Corporation (Madison, United States); triethylammonium bicarbonate (TEAB) buffer, iodoacetamide (IAM) and bovine serum albumin (BSA) were from Sigma-Aldrich Corporation (St Louis, MO, United States); Annexin V-FITC apoptosis detection kit was from Becton, Dickinson and Company (New Jersey, United States); cell cycle analysis kit was from Beyotime Biotechnology (Shanghai, China); acetonitrile (ACN), formic acid (FA) and TMT 6-plex reagent kit were from Thermo Fisher Scientific (Waltham, MA, United States); protease inhibitor cocktail was from Bimake (Houston, Texas, United States).
6
Immortalized Human Liver Cell Lines
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Human hepatic stellate cell line LX‐2 was obtained from the China Center for Type Culture Collection (CCTC, Wuhan, China). An immortalized human liver cell line MIHA was kindly provided by Prof. Ben C. B. Ko (The Hong Kong Polytechnic University, Hong Kong, China).[58 (link)
] Cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Gibco, Grand Island, NY, USA) supplemented with 10% FBS (Corning, NY, USA), and 1% penicillin‐streptomycin (MedChemExpress, NJ, USA) at 37 °C containing 5% CO2.
For the bile acid treatment experiments, LX‐2 cells were seeded in 6‐well plates, and cells were stimulated by fresh 2% FBS medium with or without individual BAs (Sigma‐Aldrich, St Louis, MO, USA) at 25, 50, 75, and 100 µм for 48 h, and DMSO were used as a vehicle control.
For co‐culture experiments, LX‐2 cells were seeded in 12‐well plates at densities of 50000 cells cm−2. MIHA cells were seeded and adhered to the porous polyester (PET) membrane surface of trans‐well inserts (0.4 µm pores, BIOFIL, Guangzhou, China) at a density of 100000 cells cm−2. Inserts containing MIHA cells were incubated overnight to adapt to the new conditions before being placed in 12‐well plates containing HSCs. The co‐culture system was incubated for 48 h in DMEM medium with 2% FBS. trans‐well inserts were then carefully removed and the LX‐2 cells in 12‐well plates were immediately collected for analysis.
] Cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Gibco, Grand Island, NY, USA) supplemented with 10% FBS (Corning, NY, USA), and 1% penicillin‐streptomycin (MedChemExpress, NJ, USA) at 37 °C containing 5% CO2.
For the bile acid treatment experiments, LX‐2 cells were seeded in 6‐well plates, and cells were stimulated by fresh 2% FBS medium with or without individual BAs (Sigma‐Aldrich, St Louis, MO, USA) at 25, 50, 75, and 100 µм for 48 h, and DMSO were used as a vehicle control.
For co‐culture experiments, LX‐2 cells were seeded in 12‐well plates at densities of 50000 cells cm−2. MIHA cells were seeded and adhered to the porous polyester (PET) membrane surface of trans‐well inserts (0.4 µm pores, BIOFIL, Guangzhou, China) at a density of 100000 cells cm−2. Inserts containing MIHA cells were incubated overnight to adapt to the new conditions before being placed in 12‐well plates containing HSCs. The co‐culture system was incubated for 48 h in DMEM medium with 2% FBS. trans‐well inserts were then carefully removed and the LX‐2 cells in 12‐well plates were immediately collected for analysis.
7
Endometrial Cancer Cell Culture and Hypoxia
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The endometrial cancer cell line Ishikawa (ISK) cells were cultured in the medium of DMEM/F12 (11320082, Gibco, Carlsbad, CA, USA), supplemented with 1 percent penicillin/ streptomycin (15140122, Gibco, Carlsbad, CA, USA), and 10 percent FBS (SV30087, HyClone). ECSC cells were cultured in DMEM/F12 media supplemented with 10 ng/mL Human Fibroblast Growth Factor Protein (FGF) (AA 10-155, Sigma-Aldrich, St. Louis, MO, USA), 20 ng/mL Human Epidermal Growth Factor Protein (EGF) (HY-P7109, MedChemExpress, Monmouth Junction, NJ, USA), 0.4% Bovine Serum Albumin (BSA) (HZB0148, Sigma-Aldrich, St. Louis, MO, USA) 20 ng/mL Insulin (HY-P73243, MedChemExpress, Monmouth Junction, NJ, USA) and 1% penicillin/ streptomycin. For hypoxia exposure, cells were placed in a modular incubator chamber (Billups-Rothenberg, San Diego, CA, USA), which was reported before [15] . In testing conditions, ECSC cells were cultured in defined media before supplemented chemicals as indicated as CoCl 2 , the HIFs activator (7646-79-9, Sigma-Aldrich, St. Louis, MO, USA), and KC7F2, the HIFs inhibitor (HY-18777, MedChemExpress, Monmouth Junction, NJ, USA) as used in the experiments.
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