SH-SY5Y human neuroblastoma cells (ATCC# CRL-2266) were cultured in Eagle’s Minimum Essential Medium (EMEM) supplemented with 10% fetal bovine serum, 3.0 mM glutamine, 50 units/mL penicillin and 50 mg/mL streptomycin in a 5.0% CO2 humidified environment at 37 °C. PANC-1 cells (ATCC# CRL-1469) were cultured in Dulbecco’s modified eagle’s medium (DMEM) – high glucose supplemented with 10% (v/v) heat inactivated fetal bovine serum, 50 units/mL penicillin and 50 mg/mL streptomycin at 37 °C in a humidified atmosphere with 5% CO2. The cells were plated at a density of 100,000 cells/well on 24-well plates in 1 ml of medium. After 24 h, cells were exposed to 30 µM protein samples. Cells in culture medium without protein and in the presence of curcumin and vanillin at the tested concentrations served as control. In the AGE experiments, before cells exposure, insulin glycated in the presence of 0.5 M D-ribose for 8 days was subjected to dialysis in sterile conditions to remove the free glycating agent89 (link).
Sh sy5y human neuroblastoma cells
Manufactured by American Type Culture Collection
Sourced in United States
The SH-SY5Y human neuroblastoma cells are a subclone of the SK-N-SH cell line derived from a human neuroblastoma. They are widely used in research as a model for neuronal cells.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (12 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (6 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (6 mentions)
Fetal bovine serum (fbs), by Merck Group (5 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (4 mentions)
Sh sy5y cells, by Thermo Fisher Scientific (4 mentions)
Penicillin, by Merck Group (3 mentions)
Streptomycin, by Merck Group (3 mentions)
L glutamine, by Merck Group (3 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions)
Penicillin, by Thermo Fisher Scientific (3 mentions)
Streptomycin, by Thermo Fisher Scientific (3 mentions)
Xtt cell viability assay kit, by Biotium (2 mentions)
Synergy htx multi mode reader, by Agilent Technologies (2 mentions)
Glutamine, by Merck Group (2 mentions)
Jurkat human t cell leukemia cells, by American Type Culture Collection (2 mentions)
Dulbecco s modified eagle s medium, by Merck Group (2 mentions)
100 mm culture dishes, by SPL Life Sciences (2 mentions)
Penicillin streptomycin amphotericin b mixture, by Thermo Fisher Scientific (2 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (2 mentions)
78 protocols using sh sy5y human neuroblastoma cells
1
In vitro neuroblastoma and pancreatic cancer cell culture
2
Dopaminergic Receptor Activation Assay
3
SH-SY5Y Cell Culture Protocol
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SH-SY5Y human neuroblastoma cells were purchased from ATCC (ref: CRL-2266). Cells were seeded at 5x104 cells/cm2 and used 5 days later, usually when cultures reached a 70–80% confluence. Culture media contained DMEM F-12 Ham (Sigma-Aldrich, USA) supplemented with 0.5 mM glutamine (Sigma-Aldrich, USA), penicillin (50 mg/ml)/streptomycin (50 U/ml) from Sigma-Aldrich (USA), and 10% FBS (Sigma-Aldrich, USA). Cells were serum starved for 16 hours prior to treatment, in order to reduce Akt basal activity.
4
Aβ Oligomer Cytotoxicity in SH‐SY5Y Cells
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SH‐SY5Y human neuroblastoma cells (ATCC, Manassas, VA) were plated in a clear 96 well‐plate at a density of approximately 20 000 cells/well in growth media containing 1:1 mixture of DMEM/F12 medium with 10% fetal bovine serum and 1% Pen‐Strep at 37°C in a humidified atmosphere of 5.5% CO2. After 24 hours, media was replaced with freshly prepared Aβ oligomers in siliconized tubes resuspended in growth media and incubated for 40 hours. Cell viability assay was performed using 2,3‐bis(2‐methoxy‐4‐nitro‐5‐sulfophenyl)‐5‐[(phenylamino)carbonyl] ‐2H‐ tetrazolium hydroxide (XTT) cell viability assay kit (Biotium). Absorbance was measured at 475 and 660 nm using Synergy HTX Multi‐Mode Reader‐ BioTek Instruments.
5
Neuroblastoma Cell Viability Assay
6
Neuroblastoma Cell Treatment and Analysis
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SH-SY5Y human neuroblastoma cells were purchased from ATCC® (Manassas, VA). Cells were grown as proliferative monolayers in 10 cm standard tissue culture dishes, containing 10 mL of alpha-modified Minimum Essential Medium (α-MEM) from Mediatech (Manassas, VA) supplemented with 1 % penicillin-streptomycin-fungizone, also from Mediatech, and 10 % fetal bovine serum (FBS) from HyClone (Logan, UT) at 37 °C with 5 % CO2. Cells (Passage # 4) treated for 4 h with 1 μM hBCM7, bBCM7, morphine or left untreated as a control prior to RNA or DNA extraction. This concentration was chosen on the basis of previous dose–response studies indicating that 1 μM produced maximum inhibition of EAAT3-mediated cysteine uptake.
7
Comprehensive Analysis of Neuronal Cytoprotection
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MC, BR and ADR were commercially purchased from Kyungdong Herbal Market (Seoul, Korea) (see Supplementary Material for details of each herb). The primary antibodies against tyrosine hydroxylase (TH), AKT, pAKT(T308), and pAKT(S473) were purchased from Cell Signaling Technology (Beverly, MA, USA). Anti-β-actin antibody was purchased from Sigma-Aldrich Co. (St. Louis, CO, USA). Horseradish peroxidase (HRP)-conjugated secondary antibodies were purchased from Cell Signaling Technology. Biotinylated anti-rabbit antibody and the avidin-biotin peroxidase complex (ABC) standard kit were purchased from Vector Laboratories (Burlingame, CA, USA). Calcein AM, tetramethylrhodamineethylester (TMRE), 5,6-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate acetyl ester (CM-H2DCFDA), and MitoSox agents were purchased from Molecular Probes (Eugene, OR, USA). SH-SY5Y human neuroblastoma cells were purchased from ATCC® (CRL-2266™; Manassas, VA, USA). All other reagents were purchased from Sigma-Aldrich. Media and culture reagents were products of Gibco Industries Inc. (Auckland, New Zealand).
8
Cell Culture and Differentiation Protocols
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K562 and TF-1 human erythroleukemia and Jurkat human T-cell leukemia cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA). They were cultured in RPMI 1640 medium containing 10% fetal bovine serum (FBS; Mediatech, Herndon, VA), 25 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, 2 mM l -glutamine, and 4.5 g/l glucose. Jurkat and TF-1 culture media contained 1 mM sodium pyruvate, and TF-1 cells were cultured in the presence of 2 ng/ml granulocyte-macrophage colony-stimulating factor. SH-SY5Y human neuroblastoma cells were also purchased from ATCC. They were cultured in 1:1 mixture of MEM with nonessential amino acids and Ham's F12 medium containing 10% FBS. They were maintained at 37°C in a humidified 95% air, 5% carbon dioxide incubator. For K562 erythroid differentiation, exponentially growing cells were exposed to hemin (Fluka-Sigma-Aldrich, St. Louis, MO). hemin and sodium arsenite (Thermo Fisher Scientific, Waltham, MA) were dissolved in 100 μM NaOH and distilled water, respectively. t-BHQ (Sigma-Aldrich, St. Louis, MO) was dissolved in dimethyl sulfoxide for 1 M solution and further diluted to 10 mM with distilled water before addition to the culture media. Sulforaphane (Sigma-Aldrich) was also dissolved in dimethyl sulfoxide for 100 mM solution.
9
Culturing SH-SY5Y Human Neuroblastoma Cells
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SH-SY5Y human neuroblastoma cells (purchased from ATCC (Rockville, MD, USA)) were cultured in DMEM/F12 media supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin at 37 °C with 5% CO2. The medium was changed every other day, and cells were plated at an appropriate density according to each experimental scale.
10
Cell Culture and Aβ Aggregation Protocol
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U373-MG human glioblastoma astrocytoma cells and SH-SY5Y human neuroblastoma cells were purchased from ATCC (Manassas, VA, USA). Cells were grown in DMEM supplemented with 2 mM L-glutamine, 10% FBS and 40 μg/mL gentamicin at 37 °C in a humidified 5% CO2 incubator. Confluent monolayers of U373 cells were sub-cultured by conventional trypsinization. For the experiments, 2.5 × 105 or 4 × 105 cells were seeded in 35 or 60 mm tissue culture dishes, respectively, and grown up to 80% confluence for 18–24 h before treatments. Stock solution of Aβ25-35 was prepared at 2.5 mM concentration in bi-distilled water and kept frozen at −20 °C. For aggregation, Aβ25–35 was aged overnight at 25 °C before being added to the culture medium to the final desired concentration (for further details on Aβ25–35 preparation, conformation and aggregation properties see [9 (link),30 (link)]).
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