The human CRC cell lines SW480, COLO320, T84 were purchased from the American Type Culture Collection (ATCC; Rockville, MD, USA). The human normal intestinal mucous cell line CCC-HIE-2 and other CRC cell lines SW620, LoVo and HCT116 were obtained from the Type Culture Collection of the Chinese Academy of Medical Sciences (Beijing, China). The cell lines were maintained at 37°C in a 5% humidified CO2 atmosphere. CCC-HIE-2 cells were cultured in DMEM supplemented with 20% FBS and 2ng/ml EGF (Solarbio, Beijing, China). HCT116, LoVo and other CRC cell lines were cultured in IMDM, F12K and RPMI-1640, respectively, with 10% FBS.
Epidermal growth factor (egf)
Manufactured by Solarbio
Sourced in China, United States
EGF is a growth factor that stimulates cell growth and differentiation. It is a protein involved in the regulation of cell proliferation, survival, and migration.
Automatically generated - may contain errors
Lab products found in correlation
Basic fibroblast growth factor (bfgf), by Solarbio (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Sw480, by American Type Culture Collection (1 mentions)
Colo320, by American Type Culture Collection (1 mentions)
Dmem high glucose culture medium, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Solarbio (1 mentions)
Ultra lowattachment 24 wells plates, by Corning (1 mentions)
Ix53 dp80, by Olympus (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Low attachment 6 well plate, by Corning (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
Neurobasal medium, by Thermo Fisher Scientific (1 mentions)
Heparin, by Solarbio (1 mentions)
Hydrocortisone, by Solarbio (1 mentions)
24 well ultra low attachment plate, by Corning (1 mentions)
Costar ultra low attachment 96 well plate, by Corning (1 mentions)
Dmem f12 medium, by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Solarbio (1 mentions)
6 protocols using epidermal growth factor (egf)
1
Characterization of CRC Cell Lines
2
Goose Embryonic Hepatocyte Stress Response
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After the fertilized goose eggs were incubated for 23 days, the embryos were sacrificed for liver tissue collection. According to the protocol described previously [45 (link)], primary hepatocytes were prepared and plated at a density of 5 × 106 cells per well. The cells were incubated overnight in a complete culture medium containing high-glucose DMEM culture medium (Gibco, New York, NY, USA), 10% fetal bovine serum (Gibco, New York, NY, USA), 1% penicillin–streptomycin (Solarbio, Beijing, China) and 0.1% EGF (Solarbio, Beijing, China). For glucose treatment, the cells in complete culture medium were treated with 0 mmol/L (the control group), 50 mmol/L and 100 mmol/L glucose, respectively. For oleate treatment, the cells in complete culture medium were treated with 0 mmol/L (the control group), 0.125 mmol/L and 0.25 mmol/L sodium oleate, respectively. For thyroxine treatment, the cells in complete culture medium were treated with 0 µmol/L (the control group), 0.1 µmol/L and 0.3 µmol/L sodium thyroxine, respectively. After 14 h of treatment, the cells were collected for later analysis.
3
Ovarian Cancer Tumorsphere Cultivation
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Single-cell suspensions of ovarian cancer cells (4 × 102 cells/mL) were plated on ultra-low attachment 24 wells plates (Corning, America) and cultured in phenol red-free DMEM/F12 (Gibco, America) containing B27 supplement (Gibico, America, #12587010) and 20 ng/mL epidermal growth factor (EGF, Solarbio, China, #P00033), 20 ng/mL basic fibroblast growth factor (bFGF, Solarbio, China, #P00032), and 5 μg/mL insulin (Solarbio, China, #I8040). Tumorsphere were visualized under a phase-contrast microscope (Olympus, America, IX53 + DP80).
4
Triple-Negative Breast Cancer Spheroid Assay
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Human triple-negative breast cancer cell line MDA-MB-231 and mouse triple-negative breast cancer cell line 4T1 were purchased from ATCC and cultured in DMEM medium with 10% fetal bovine serum and was aired with 5% CO2 at 37 °C. Single MDA-MB-231 cells or 4T1 cells were plated in low attachment 6-well plates (Corning, Oneonta, NY, USA) at a density of 5000 cells/well, and cultured in serum-free DMEM/F12k containing B27 (Thermo Fisher Scientific, Inc., Waltham, MA, USA), epidermal growth factor (EGF, 20 ng/mL; Solarbio Science and Technology Co., Ltd., Beijing, China), and basic fibroblast growth factor (bFGF20 ng/mL, Solarbio Science and Technology Co., Ltd., Beijing, China). The cells were allowed to grow for 7 days; then, the medium was changed, and they were cultured for another 7 days. The quantification of the number of spheres was carried out using microscopy to count all spheres with a diameter bigger than 50 μm. Experiments were repeated three times, with three replicates each.
5
Culturing PC12 Cells and Isolating Rat NSCs
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Rat pheochromocytoma (PC12) cells (CL-0412, Wuhan Procell Co., Ltd) were cultured in RPMI-1640 medium (Gibco) with 5% fetal bovine serum (FBS, Gibco), 10% horse serum (HS, Gibco) and 1% penicillin/streptomycin (PS, Gibco). Neural stem cells (NSCs) were isolated from the hippocampus of E13 Sprague-Dawley rat embryos as described previously [1 (link)]. Briefly, the hippocampus was gently extracted, stripped of blood vessels, cut into small pieces, and digested with 0.05% EDTA/trypsin (Gibco, USA) at 37 °C for 10 min, followed by centrifugation at 200×g for 5 min. Cells were cultured in proliferative NSC medium containing Neurobasal medium (Gibco, USA), 2% B27 (Gibco, USA), 1% PS (Sigma-Aldrich, USA), 1% glutamax (Gibco, USA), 20 ng/mL basic fibroblast growth factor (bFGF, Solarbio, China), and 20 ng/mL epidermal growth factor (EGF, Solarbio, China) in a 5% CO2 humidified 37 °C incubator. For further experiment, 1 × 105 PC12 cells or 1 × 106 were seeded on each sample, respectively. To make sure the cell adhering on the hydrogel surface, the 1 mL cell suspension was dropped onto the hydrogel surface, and after 30 min later the cell culture medium was gently injected into the well from the sidewall to immerse the hydrogel fibers.
6
Mammosphere Formation Assay for Breast Cancer Stem Cells
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Single cells were seeded into the ultralow attachment 24‐well plate (Corning, NY, USA) at a density of 2 × 103 per well with serum‐free DMEM/F‐12 medium (Gibco) supplemented with 1 × B27 (Gibco), 0.4% bovine serum albumin (Solarbio), 0.5 µg mL−1 hydrocortisone (Solarbio), 4 µg mL−1 heparin (Solarbio), 4 µg mL−1 insulin (Solarbio), 20 ng mL−1 EGF (Solarbio), and 20 ng mL−1 bFGF (Solarbio). Cells were cultured for 5–7 days with 500 µL fresh medium added every 3 days, and spheres (> 50 µm) were then counted and photographed using a light field microscope equipped with a phase‐contrast module (Leica, Wetzlar, Germany). Mammosphere formation efficiency (MFE) was calculated as follows: MFE (%) = (the number of mammospheres identified per well/the number of cells seeded per well) × 100%. For the limiting dilution analysis of the mammosphere formation, single cells were seeded into the Corning Costar ultralow attachment 96‐well plate at the density of 1–2 × 102 per well. After 5–7 days, the percentage of wells without spheres was plotted against the number of cells per well, and the regression lines were generated accordingly.
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