AsPC-1, BxPC-3, MiaPaCa-2, CFPAC-1, PANC1 and hTERT-HPNE cell lines were from ATCC® (Manassas, VA, USA). The murine Kras-driven pancreatic cancer cell line MT4-2D was kindly provided by Professor David. Tuveson (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, USA). All cells tested negative for Mycoplasma. AsPC-1 and BxPC-3 cells were maintained in RPMI-1640 medium. MiaPaCa-2, PANC-1 and MT4-2D were maintained in DMEM medium. CFPAC-1 cells were maintained in IMDM medium. All culture media were supplemented with 10% fetal bovine serum (FBS, from Bovogen Biologicals, Melbourne, Australia), 2 mM glutamine, 1 mM sodium pyruvate and 1 mM non-essential amino acids. hTERT-HPNE cells were maintained in DMEM medium supplemented with 5% FBS, human epidermal growth factor (EGF 10 ng/mL, ThermoFischer, Waltham, MA, USA), puromycin (750 ng/mL, ThermoFischer, Waltham, MA, USA) and 5 mM D-glucose. Unless specified, all reagents were obtained from Gibco® Life Technologies (Melbourne, Australia).
Human epidermal growth factor egf
Manufactured by Thermo Fisher Scientific
Sourced in United States
Human epidermal growth factor (EGF) is a protein that stimulates cell growth, proliferation, and differentiation. It is a key regulator of various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Bovine insulin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Mia paca 2, by American Type Culture Collection (1 mentions)
Bxpc 3, by American Type Culture Collection (1 mentions)
Puromycin, by Thermo Fisher Scientific (1 mentions)
Panc 1, by American Type Culture Collection (1 mentions)
Aspc 1, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Bovogen (1 mentions)
Htert hpne, by American Type Culture Collection (1 mentions)
Cfpac 1, by American Type Culture Collection (1 mentions)
Gentamicin amphotericin, by Thermo Fisher Scientific (1 mentions)
Trypsin edta solution, by Thermo Fisher Scientific (1 mentions)
Human keratinocyte growth supplement, by Thermo Fisher Scientific (1 mentions)
Hydrocortisone, by Thermo Fisher Scientific (1 mentions)
Collagenase, by Merck Group (1 mentions)
Dispase, by Thermo Fisher Scientific (1 mentions)
Bovine transferrin, by Thermo Fisher Scientific (1 mentions)
Epilifecf, by Thermo Fisher Scientific (1 mentions)
Human transferrin, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
6 protocols using human epidermal growth factor egf
1
Pancreatic Cancer Cell Line Culture
2
Primary Keratinocyte Isolation from Cervix
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Primary Keratinocytes (Kers) were obtained from the normal cervixes of women who underwent hysterectomy, and the samples were taken under signed informed consent at the Western Medical Center -IMSS. Biopsies were collected in EPILIFE CF medium (Thermo Fisher Scientific, Inc.) with 100 U/ml Penicillin and 100 μg/ ml Streptomycin. After removal of connective and fatty tissue, the biopsy was chopped into 0.5-cm2 fragments and treated with 25 U/ml collagenase (Sigma-Aldrich Quimica, S. A. de R.L. de C.V, Toluca, México) and 25 U/ml dispase (Thermo Fisher Scientific, Inc.) overnight at 4oC with constant movement. Then, the epidermis was removed from connective tissue with tweezers. Epithelial cells were disrupted by incubation for 30 min at 37 o C in 2 ml of 0.25% trypsin-EDTA solution (Thermo Fisher Scientific, Inc.). Trypsin was neutralized with FBS and the cells were collected by centrifugation and resuspended in 13 ml of selective medium for keratinocytes EPILIFE CF supplemented with Human Keratinocyte Growth Supplement containing: 0.2% v/v pituitary gland extract (BPE), 5 μg/ml bovine insulin, 0.18 μg/ml hydrocortisone, 5 μg/ml bovine transferrin, 0.2 ng/ml human Epidermal Growth Factor (EGF), and Gentamicin/Amphotericin (all from Thermo Fisher Scientific, Inc.). The cultures were maintained at 37 o C in a humidified atmosphere with 5% CO 2 .
3
Culturing Human Corneal Epithelial Cells
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In this study, HCECs were provided by Ophthalmology and Optometry and Eye Hospital in Wenzhou, China. The cells were incubated under 5% CO2 at 37 °C, and cultured in DMEM (Waltham, MA, USA) with high glucose, supplemented with 15% fetal bovine serum (Gibco BRL), 5 μg mL−1 insulin, 100 U mL−1 penicillin, 5 μg mL−1 human transferrin (Sigma-Aldrich, MA, USA), 2 mM L-glutamine, 100 μg mL−1 streptomycin (HyClone) and 10 ng mL−1 human epidermal growth factor (EGF; Gibco BRL). The Col-Tau sample was washed three times in PBS and sterilized by UV radiation for about 2 h before cell experiments. Then, the pre-wetted sterile sample was used for cell culture and the seeded density of HCECs was 5000 cells/cm2. During cell culture, the cell culture medium was changed every two days. The change of HCECs on the surface of film was observed under an inverted phase contrast microscope (Zeiss Observer A1). Moreover, the metabolic activity of the HCECs was determined by MTT assay.
4
Detailed Cell Culture Protocol
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SAOS-2 cells were cultured in McCoys 5A medium (modified) (Sigma-Aldrich) supplemented with 2.2 g/l sodium bicarbonate, 10% heat inactivated foetal calf serum (FCS) and 1 mM sodium pyruvate. SR5 and SR40 cells were cultured in the above medium with an additional 800 μg/ml Geneticin (GIBCO) as a selective agent. MCF10A, MCF10AT and MCF10CA1h cells were cultured in DMEM/F12 medium with 10 mM HEPES (Sigma), supplemented with 2.2 g/l sodium bicarbonate, 5% horse serum, 100 ng/ml cholera toxin (Sigma), 0.5 μg/ml hydrocortisone, 10 μg/ml bovine insulin and 20 ng/ml human epidermal growth factor (EGF) (GIBCO). All cells were grown at 37°C in 5% CO2. Cells were seeded onto glass coverslips (13 mm in diameter or 15 X 15 mm) 24 h prior to transduction experiments or transfection conducted using 4 μg of DNA complexed with 10 μl LipofectAMINE 2000™ (Invitrogen) per well of a 6-well tissue culture plate, according to the manufacturer’s instructions.
5
Genotoxicity Assessment of Chemicals and NPs
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Immortalized FE1 cells derived from the Muta™ Mouse transgenic rodent model were utilized for particle and metal chloride exposures. These cells retain the characteristics of type I and type II pulmonary alveolar epithelial cells, and have been used in the past to assess the genotoxicity and mutagenicity of both chemicals and NPs [30 (link),31 (link),32 (link),33 (link),34 (link),35 (link)].
Cells were maintained in phenol-red-containing Dulbecco’s Modified Eagle’s Medium Nutrient Mixture: F12 HAM (1:1) culture media (DMEM/F12 (1:1) (1X), Life Technologies, Burlington, ON, Canada) supplemented with 2% fetal bovine serum (FBS, Life Technologies, Burlington, ON, Canada), 1 ng/mL human epidermal growth factor (EGF, Life Technologies, Burlington, ON, Canada), 1 U/mL penicillin G, and 1 µg/mL streptomycin (Life Technologies, Burlington, ON, Canada) in an incubator at 37 °C, with 5% CO2. For CometChip exposures, the same conditions were used, except for the absence of phenol-red in the exposure media.
Cells were maintained in phenol-red-containing Dulbecco’s Modified Eagle’s Medium Nutrient Mixture: F12 HAM (1:1) culture media (DMEM/F12 (1:1) (1X), Life Technologies, Burlington, ON, Canada) supplemented with 2% fetal bovine serum (FBS, Life Technologies, Burlington, ON, Canada), 1 ng/mL human epidermal growth factor (EGF, Life Technologies, Burlington, ON, Canada), 1 U/mL penicillin G, and 1 µg/mL streptomycin (Life Technologies, Burlington, ON, Canada) in an incubator at 37 °C, with 5% CO2. For CometChip exposures, the same conditions were used, except for the absence of phenol-red in the exposure media.
6
Mammosphere Formation Assay for Cancer Stem Cells
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MDAMB231, MCF7-sh-scrambled and MCF7-sh-WISP2 cell were plated in ultra-low attachment plate (Nunc®) in medium containing DMEM/F12 serum-free, supplemented with 100µg/ml gentamycin (Sigma-Aldrich), B27 (Life Technologies), 20 ng/ml human epidermal growth factor (EGF, Life Technologies), 20 ng/ml Human basic fibroblast growth factor (bFGF, Life Technologies) and 1% antibiotic-antimycotic solution (Life Technologies) at a density of 20,000 cells/ml. Mammosphere medium was added to plates every four days. After two weeks, they were collected, dissociated in 0.05% trypsin, 0.25% EDTA and re-seeded at 10.000 cells/ml for second passages. Enumeration of mammospheres was realized for first and second generations. The mammosphere-containing media was harvested, gently centrifuged, rinsed carefully not to disturb the sphere pellets, and re-suspended in the appropriate medium. The sphere suspensions were counted on a microscope Nikon Eclipse at X10. Results are expressed as a percentage of mammosphere forming units (%MFU) from the total number of cells plated.
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