Trypsin edta

Manufactured by Euroclone

Sourced in Italy, United States

Trypsin-EDTA is a solution used for the dissociation and detachment of adherent cells in cell culture. It contains the enzyme trypsin, which cleaves peptide bonds, and EDTA, which chelates calcium ions to disrupt cell-cell and cell-substrate adhesion. This product is commonly used to passage or subculture adherent cells.

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Lab products found in correlation

Penicillin, by Euroclone (14 mentions) L glutamine, by Euroclone (14 mentions) Streptomycin, by Euroclone (13 mentions) Fetal bovine serum (fbs), by Euroclone (11 mentions) Penicillin streptomycin, by Euroclone (6 mentions) Phosphate buffered saline (pbs), by Euroclone (6 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions) Bovine serum albumin (bsa), by Merck Group (5 mentions) Glutamine, by Euroclone (5 mentions) Triton x 100, by Merck Group (4 mentions) Petri dishes, by Euroclone (4 mentions) Glutamax, by Thermo Fisher Scientific (3 mentions) Hydrogen peroxide h2o2, by Merck Group (3 mentions) Sodium pyruvate, by Euroclone (3 mentions) Dmem glutamax, by Thermo Fisher Scientific (3 mentions) Fetal calf serum (fcs), by Euroclone (3 mentions) Hydrocortisone hemisuccinate, by Merck Group (2 mentions) William s e medium, by Thermo Fisher Scientific (2 mentions) Heparg human progenitor hepatic cells, by Thermo Fisher Scientific (2 mentions) Penicillin, by Merck Group (2 mentions)

52 protocols using trypsin edta

Isolation and Characterization of Human Mesenchymal Stromal Cells

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Bone marrow samples were obtained from healthy donors after written informed consent. Cells were separated on Ficoll density gradient (GE Healthcare, Italy), and the mononuclear cell fraction was collected and washed in PBS. We seeded 1 to 2.5 × 10⁵ cells/cm² in alpha-minimum essential medium (alpha-MEM) (EuroClone, Italy) containing 10% fetal bovine serum (FBS) (EuroClone, Italy), 100U/mL penicillin and 100 mg/mL streptomycin (EuroClone, Italy), 1× L-Glutamine (EuroClone, Italy), and 3 ng/mL basic fibroblast growth factor (bFGF) (Preprotech, United Kingdom). After 72 h, non-adherent cells were discarded and adherent cells were further cultivated to confluency and amplified at P1. Then, the medium was changed every 3 d and cells harvested when they reached 70%–80% confluence using Trypsin-EDTA (EuroClone, Italy) and routinely sub-cultured at 1:3 dilution.
We evaluated on MSCs the expression of markers recognized as one of the criteria to identify MSCs[20 (link)]. We verified by immunocytochemistry that MSCs expressed the surface antigens CD73, CD90 and CD105.

Alessio N., Squillaro T., Monda V., Peluso G., Monda M., Melone M.A., Galderisi U, & Di Bernardo G. (2019). Circulating factors present in the sera of naturally skinny people may influence cell commitment and adipocyte differentiation of mesenchymal stromal cells. World Journal of Stem Cells, 11(3), 180-195.

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Culturing Human Hepatic Cell Lines

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HepG2 human hepatocarcinoma cells were maintained in MEM medium (Gibco, Life Technology) supplemented with 10% fetal bovine serum (Gibco, Life Technology) in the presence of 100 U/ml penicillin and 100 mg/ml streptomycin (Sigma-Aldrich, USA). HepaRG human progenitor hepatic cells (Thermo Fisher Scientific) were maintained according to the manufacturer’s recommendations. Briefly, the cells were grown in William’s E medium (Thermo Fisher Scientific) supplemented with Glutamax (Gibco), 5 μg/mL human insulin (Sigma-Aldrich) and 50 μM hydrocortisone hemisuccinate (Sigma-Aldrich) for 14 days. All cells were kept at 37°C in a humidified 5% CO₂ cell culture incubator and were passaged using trypsin-EDTA (Euroclone).

Bartolini D., Wang Y., Zhang J., Giustarini D., Rossi R., Wang G.Y., Torquato P., Townsend D.M., Tew K.D, & Galli F. (2019). A seleno-hormetine protects bone marrow hematopoietic cells against ionizing radiation-induced toxicities. PLoS ONE, 14(4), e0205626.

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Antioxidant Effects of Andrographis and Curcumin

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Andrographis paniculata (95% andrographolide) and Curcuma Longa, (96.8% curcumin) were provided by Nuova Farmaceutica S.r.l. (Catania, Italy). Fetal bovine serum (FBS), penicillin, streptomycin, L-glutamine, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), and trypsin-EDTA were purchased from Euroclone S.p.A. (Pero, Italy). Isopropanol, ethanol, phosphate buffer saline (PBS), formaldehyde, and bovine serum albumin (BSA) were obtained from Sigma-Aldrich (Milan, Italy). Palmitic acid and oleic acid were bought from Starlab S.r.l. (Milano, Italy).

Pipitone R.M., Zito R., Lupo G., Javed A., La Mantia C., Di Maria G., Pratelli G., Di Salvo F., Fontana S., Pucci M., Carlisi D, & Grimaudo S. (2023). Curcumin and Andrographolide Co-Administration Safely Prevent Steatosis Induction and ROS Production in HepG2 Cell Line. Molecules, 28(3), 1261.

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Culturing PC3 Prostate Cancer Cells

Cited 1 time

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Human prostate cancer PC3 cells, derived from bone metastasis of a grade IV prostatic adenocarcinoma, were grown in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS), 100,000 units/L penicillin, 50 mg/L streptomycin and 200 mM glutamine (Euroclone, Milan, Italy). All cells were grown at 37 °C in a humidified atmosphere of 5% CO₂ in the air and routinely split by trypsinization with Trypsin-EDTA (Euroclone). Experiments were performed with base viability >98%.
The protocol used in this study followed CRAC’s design as previously described [7 (link)].

Petrella G., Corsi F., Ciufolini G., Germini S., Capradossi F., Pelliccia A., Torino F., Ghibelli L, & Cicero D.O. (2022). Metabolic Reprogramming of Castration-Resistant Prostate Cancer Cells as a Response to Chemotherapy. Metabolites, 13(1), 65.

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Culture and Maintenance of Prostate Cell Lines

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Human prostate cancer PC-3 and LNCaP cells (ATCC) were grown in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS), 100,000 units/L penicillin, 50 mg/L streptomycin and 200 mM glutamine (Euroclone). Non-neoplastic, immortalized human prostatic epithelial cells, RWPE-1 (ATCC), were grown in keratinocyte serum-free medium (K-SFM), supplemented with 1% penicillin/streptomycin (100 IU/mL), 50 μg/mL bovine pituitary extract and 5 ng/mL epidermal growth factor (Life Technologies). All cells were grown at 37 °C in a humidified atmosphere of 5% CO₂ in air and routinely split by trypsinization with Trypsin-EDTA (Euroclone). Experiments are performed with a base-viability >98%.

Corsi F., Capradossi F., Pelliccia A., Briganti S., Bruni E., Traversa E., Torino F., Reichle A, & Ghibelli L. (2022). Apoptosis as Driver of Therapy-Induced Cancer Repopulation and Acquired Cell-Resistance (CRAC): A Simple In Vitro Model of Phoenix Rising in Prostate Cancer. International Journal of Molecular Sciences, 23(3), 1152.

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Endothelial Cell Culture and Signaling

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HEPES, human endothelial serum-free medium (HE-SFM), Fast DiI, Phalloidin-Alexa Fluor 546, Trizol Reagent, Superscript II, MMLV reverse transcriptase and protease inhibitor cocktail were from Life Technologies (Carlsbad, CA, USA); Medium 199 (M199), foetal bovine serum (FBS), heparin, forskolin, LPS, Tricaine, phytohemoagglutinin (PHA) and 1-phenyl-2-thiourea were from Sigma-Aldrich (Saint. Louis, MO, USA); L-glutamine, trypsin-EDTA, penicillin-streptomycin, were from Euroclone (Siziano, IT). Collagenase was from Worthington Biochemical Corporation (Lakewood, NJ, USA). Fibronectin was from Roche (Basilea, Switzerland). Matrigel was from Becton, Dickinson & Company (Frankin Lakes, NJ, USA). Goat anti-human IL-8 blocking antibody was from Abcam (Cambridge, UK); human recombinant VEGF-A was from Immunological Sciences (Rome, IT). Monoclonal antibody anti anti-actin (clone C4), KG-501 and QNZ were from Millipore (Billerica, MA, USA); rabbit polyclonal antibodies anti-phospho-CREB (ser133) and anti-phospho-p65 were from Cell Signalling Technology (Danvers, MA, USA).

Pozzobon T., Facchinello N., Bossi F., Capitani N., Benagiano M., Di Benedetto G., Zennaro C., West N., Codolo G., Bernardini M., Baldari C.T., D’Elios M.M., Pellegrini L., Argenton F, & de Bernard M. (2016). Treponema pallidum (syphilis) antigen TpF1 induces angiogenesis through the activation of the IL-8 pathway. Scientific Reports, 6, 18785.

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Fabrication of PCL-based Biomaterials

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PCL pellets (50 kDa) were purchased from Cellink AB (Gothenburg, Sweden). The antibiotics, culture media, and trypsin-EDTA used for cell culture were purchased from Euroclone (Milan, Italy). Human platelet lysate (PL) was purchased from Carlo Erba reagents (Milan, Italy). Mannitol, Nile red, sodium alginate, calcium chloride, and protamine were from Sigma Aldrich (Milan, Italy).

Bari E., Scocozza F., Perteghella S., Segale L., Sorlini M., Auricchio F., Conti M, & Torre M.L. (2022). Three-Dimensional Bioprinted Controlled Release Scaffold Containing Mesenchymal Stem/Stromal Lyosecretome for Bone Regeneration: Sterile Manufacturing and In Vitro Biological Efficacy. Biomedicines, 10(5), 1063.

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Cell Culture of Prostate Cancer Lines

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Human mHSPC LNCaP and mCRPC PC3 cells (ATCC, Rockville, MD, USA) were grown in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS), 100,000 units/L penicillin, 50 mg/L streptomycin and 200 mM glutamine (Euroclone, Milan, Italy), at 37 °C in a humidified atmosphere of 5% CO₂ in air and routinely split by trypsinization with Trypsin-EDTA (Euroclone, Milan, Italy).
For all experiments, unless otherwise specified, LNCaP and PC-3 cells were seeded at a cell density of 17,500/cm² and 12,500/cm², respectively. Experiments are performed with a base-viability >98%.
Viable cell number was evaluated at selected time points following trypsinization and was assessed using a Burker counting chamber by the trypan blue-exclusion method.

Pelliccia A., Capradossi F., Corsi F., Tarquini G.D., Bruni E., Reichle A., Torino F, & Ghibelli L. (2023). Androgen Deprivation Freezes Hormone-Sensitive Prostate Cancer Cells in a Reversible, Genetically Unstable Quasi-Apoptotic State, Bursting into Full Apoptosis upon Poly(ADP-ribose) Polymerase Inhibition. International Journal of Molecular Sciences, 24(3), 2040.

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Hexarelin-Induced Oxidative Stress Assay

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Hexarelin, Dulbecco’s Modified Eagle’s Medium (DMEM)-high glucose, hydrogen peroxide (H₂O₂), 3 (4,5 dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT), Griess reagent, poly-D-lysin hydrobromide, 4’,6-diamidino-2-phenylindole dihydrochloride (DAPI), fluoromount aqueous mounting medium and bovine serum albumin (BSA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Penicillin, streptomycin, L-glutamine, trypsin-EDTA, phosphate-buffer saline (PBS) and fetal bovine serum (FBS) were obtained from Euroclone (Pero, Milan, Italy). Alexa Fluor 488 Phalloidin was purchased by ThermoFisher Scientific (Waltham, MA, USA). Prior to assay, Hexarelin was dissolved in ultrapure water; both Hexarelin and H₂O₂ were diluted in culture medium to final working concentrations.

Meanti R., Rizzi L., Bresciani E., Molteni L., Locatelli V., Coco S., Omeljaniuk R.J, & Torsello A. (2021). Hexarelin Modulation of MAPK and PI3K/Akt Pathways in Neuro-2A Cells Inhibits Hydrogen Peroxide—Induced Apoptotic Toxicity. Pharmaceuticals, 14(5), 444.

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ARPE-19 Cell Culture and Glucose Conditions

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The human cell line ARPE-19 (American Type Culture Collection, Manassas, VA, USA) from passages 22 to 28 were grown in DMEM/F12 1:1 medium (Euroclone, Milan, Italy) supplemented with 10% fetal bovine serum and 2 mmol/L glutamine (Euroclone, Milan, Italy) at 37 °C in 5% CO₂. The cell medium was replaced every 2 days. Cells were grown to confluence, removed with trypsin-EDTA (Euroclone, Milan, Italy), and then seeded in multiwell plates for all experiments. Before each experiment, cells were washed with phosphate-buffered saline (PBS) (Euroclone, Milan, Italy) and cultured under normal glucose conditions (1 g/L glucose, CTR). Once they reached confluence, cells were cultured in control medium (CTR) or high glucose (4.5 g/L glucose, HG) for 24 h and processed for each analysis.

Puddu A., Ravera S., Panfoli I., Bertola N, & Maggi D. (2022). High Glucose Impairs Expression and Activation of MerTK in ARPE-19 Cells. International Journal of Molecular Sciences, 23(3), 1144.

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