Human neuroblastoma SH-SY5Y cells (catalog number #94030304 European Collection of Cell Cultures) were cultured in DMEM-High glucose medium supplemented with 15% Foetal Bovine Serum (FBS), 2 mM L-glutamine and 1% non-essential amino acids (all from Euroclone), at 37°C in humified atmosphere with 5% CO2, as described (Pezzini et al., 2017 (link)).
Sh sy5y cells
Manufactured by European Collection of Cell Cultures
Sourced in United Kingdom
SH-SY5Y cells are a subclone of the SK-N-SH cell line, which was originally derived from a metastatic bone tumor biopsy of a human neuroblastoma. These cells are commonly used in research as a model for neuronal cells.
Automatically generated - may contain errors
Lab products found in correlation
Non essential amino acids (neaa), by Euroclone (1 mentions)
Foetal bovine serum (fbs), by Euroclone (1 mentions)
L glutamine, by Euroclone (1 mentions)
Dulbecco s modified eagle s medium dmem, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by European Collection of Cell Cultures (1 mentions)
Hoechst 33342, by Thermo Fisher Scientific (1 mentions)
Clarity max western ecl substrate, by Bio-Rad (1 mentions)
Nifedipine, by Merck Group (1 mentions)
ω conotoxin mviic, by Bio-Techne (1 mentions)
5 dodecanoylaminofluorescein di β d galactopyranoside c12fdg, by Thermo Fisher Scientific (1 mentions)
Fura 2 acetoxymethyl ester fura 2 am, by Merck Group (1 mentions)
Ml 218, by Bio-Techne (1 mentions)
Thapsigargin tg, by Abcam (1 mentions)
Hela cells, by Leibniz Institute DSMZ (1 mentions)
10 protocols using sh sy5y cells
1
Culturing Human Neuroblastoma SH-SY5Y Cells
2
SH-SY5Y Exposure to Amyloid and Alpha-Synuclein
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SH-SY5Y cells (European Collection of Cell Cultures, ECACC) were exposed to each sample condition; control (no protein added), Aβ40, Aβ42 and asyn (monomer, oligomer and fibril) in T25 flasks at ~80% confluence and incubated for 24 h prior to harvest via trypsinisation. Samples were centrifuged for 3 min at 300 g, media was discarded and cells were washed twice in 1 mL phosphate-buffered saline and centrifuged again. Cell pellets were stored at − 80 °C. All conditions were tested in three biological replicates, and each biological replicate provided two technical replicates of ~150,000 cells per sample. Cells were assessed for viability prior to freezing using trypan blue (Strober 2001 (link)).
3
Culturing Human Neuroblastoma Cells
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Human neuroblastoma SH-SY5Y cells were obtained from ECACC (European Collection of Cell Cultures, Salisbury, UK) and cultured according to its recommendations.
4
SH-SY5Y Cell Culture Protocol
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SH-SY5Y cells were obtained from the European Collection of Cell Cultures (Porton Down, UK), Dulbecco’s Modified Eagle’s Medium (DMEM) from Sigma-Aldrich (St. Louis, MO, United States), and GA from Nacalai Tesque, Inc. (Kyoto, Japan). All other chemicals were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). The cultivation of SH-SY5Y cells was performed in DMEM supplemented with 10% fetal bovine serum, 100 U/mL of penicillin, and 100 μg/ml of streptomycin at 37°C in a humidified atmosphere of 95% air and 5% CO2.
5
SH-SY5Y Neuroblastoma Cell Culture
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Human neuroblastoma SH-SY5Y cells from the European Collection of Cell Cultures (ECACC No. 94030304) were cultured in a medium with equal amounts of Eagle’s minimum essential medium and Nutrient Mixture Ham’s F-12, supplemented with 10% heat-inactivated fetal bovine serum (FBS), 2 mM glutamine, 0.1 mg/ml streptomycin, 100 IU·ml penicillin and non-essential amino acids at 37°C in 5% CO2-containing, and 95% air atmosphere. All culture media, supplements and FBS were purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany).
6
SH-SY5Y Cell Culture and Characterization
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SH-SY5Y cells were purchased from European Collection of Cell Cultures (ECACC) and distributed by Sigma-Aldrich (St. Louis, MO, USA); Phoenix-AMPHO HEK293 were from American Type Culture Collection (ATCC); all-trans-retinoic acid, human brain-derived neurotrophic factor (BDNF), collagen (type I), Dulbecco’s modified Eagle’s medium (DMEM), DMEM:F-12 Ham medium, and nifedipine were purchased from Sigma-Aldrich (St. Louis, MO, USA); fura-2-acetoxymethyl ester (fura-2-AM) and BTP2 were from Merck Millipore (Darmstadt, Germany); thapsigargin (Tg) was from Abcam Biochemicals (Cambridge, UK); ML 218 and ω-conotoxin MVIIC were from Tocris Bioscience (Bristol, UK); rhodamine 123, tetramethylrhodamine methyl ester (TMRM), Hoechst 33258, Hoechst 33342, and 5-dodecanoylaminofluorescein di-β-d -galactopyranoside (C12FDG) were from Thermo Fisher Scientific (Waltham, MA, USA); Clarity Max™ Western ECL substrate was from Bio-Rad (Hercules, CA, USA).
7
Cortisol-Induced Stress Response in Neuroblastoma
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Human neuroblastoma SH-SY5Y cells from the European Collection of Cell Cultures (ECACC No. 94030304) were cultured in a medium with equal amounts of Eagle's minimum essential medium and Nutrient Mixture Ham's F-12, supplemented with 10 % heat-inactivated fetal bovine serum (FBS), 2 mM glutamine, 0.1 mg/ml streptomycin, 100 IU/ml penicillin and 1X MEM non-essential amino acid solution (M7145, Merck) at 37 °C in 5 % CO2-containing, and 95 % air atmosphere. SH-SY5Y cells were treated for different times with increasing concentrations of cortisol (0.1, 0.5, 1 and 5 μM) according to previous data (Buoso et al., 2011 (link); Del Vecchio et al., 2009 (link)). To demonstrate the role of glucocorticoid receptor, cells were treated for 1 h with 10 μM mifepristone before the addition of cortisol for 24 h. Other specific details of times and concentrations are also given in figure legends.
8
Culturing Neuroblastoma Cell Lines
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Human neuroblastoma SH-SY5Y cells (The European Collection of Cell Cultures) were grown in Roswell Park Memorial Institute medium with 15% FBS and 2 mM L-glutamine and kept at 37℃/5% CO2; 0.25% trypsin/EDTA was used to passage the cells every three to five days at a 1:5 dilution ratio or when approximately 90% confluent.
Mouse neuroblastoma/rat embryonic DRG neuron hybrid F-11 cells (The European Collection of Cell Cultures) were grown in Ham’s F12 media with 10% FBS, 100 mM hypoxanthine, 0.4 mM aminopterin, and 16 mM thymidine (HAT media supplement Hybri-Max™) and kept at 37℃/5% CO2; 0.25% trypsin/EDTA was used to passage the cells every two to three days at a 1:5 dilution ratio or when approximately 80% confluent.
Mouse neuroblastoma/rat DRG neuron hybrid ND7/23 cells (The European Collection of Cell Cultures) were grown in DMEM with 10% FBS, 2 mM L-glutamine, pyridoxine, and 110 mg/ml sodium pyruvate and kept at 37℃/5% CO2; 0.25% trypsin/EDTA was used to passage the cells every three to five days at a 1:5 dilution ratio or when approximately 90% confluent.
Mouse neuroblastoma/rat embryonic DRG neuron hybrid F-11 cells (The European Collection of Cell Cultures) were grown in Ham’s F12 media with 10% FBS, 100 mM hypoxanthine, 0.4 mM aminopterin, and 16 mM thymidine (HAT media supplement Hybri-Max™) and kept at 37℃/5% CO2; 0.25% trypsin/EDTA was used to passage the cells every two to three days at a 1:5 dilution ratio or when approximately 80% confluent.
Mouse neuroblastoma/rat DRG neuron hybrid ND7/23 cells (The European Collection of Cell Cultures) were grown in DMEM with 10% FBS, 2 mM L-glutamine, pyridoxine, and 110 mg/ml sodium pyruvate and kept at 37℃/5% CO2; 0.25% trypsin/EDTA was used to passage the cells every three to five days at a 1:5 dilution ratio or when approximately 90% confluent.
9
SH-SY5Y Cell Culture Protocol
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Human neuroblastoma SH-SY5Y cells were purchased from the European Collection of Cell Cultures (Salisbury, United Kingdom) and grown in Dulbecco’s modified Eagle’s F12 medium supplemented with 10% (vol/vol) heat-inactivated fetal bovine serum, 100 U/mL penicillin, and 100 µg/mL streptomycin, in a humidified atmosphere with 5% (vol/vol) CO2 in air.
10
Cultivation and Isolation of Cell Lines
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SH-SY5Y cells (European Collection of Cell Cultures; Sigma-Aldrich) were cultivated in RPMI 1640 medium containing 2 g/l glucose and 5% or 10% FCS. HeLa cells (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH) were cultivated in Minimal Essential Medium with Earle's salts containing 10% FCS and 1% non essential amino acids. Primary cortical neurons were isolated from 1–4 d old mice as described previously [3] (link) and incubated with Neurobasal medium for 10–20 days before performing experiments. All cells were kept at 37°C with 5% CO2 and 95% air.
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