Lymphocytes derived from a normal subject (HG 00866) and a G6PD-deficient subject carrying the Canton variant (HG 02367) were purchased from Coriell Institute and cultured in RPMI 1640 supplemented with 15% fetal bovine serum (FBS), 100 U mL−1 penicillin, and 100 μg mL−1 streptomycin. An SH-SY5Y neuroblastoma cell line (ATCC CRL-2266) was cultured in Dulbecco’s Modification of Eagle’s Medium/Ham’s F-12 50/50 Mix supplemented with 10% FBS, 100 U mL−1 penicillin, and 100 μg mL−1 streptomycin. A fibroblast cell line derived from a G6PD-deficient subject carrying the Mediterranean variant and normal fibroblast cell line as control were purchased from Coriell Institute (GM 01152) and Thermo Fisher Scientific (C0135C), respectively, and cultured in minimum essential medium supplemented with 15% FBS, 100 U mL−1 penicillin, and 100 μg mL−1 streptomycin. All the cell lines were maintained at 37 °C in a humidified incubator with an atmosphere of 5% of CO2 and 95% air.
Sh sy5y neuroblastoma cell line
Manufactured by American Type Culture Collection
Sourced in United States
The SH-SY5Y neuroblastoma cell line is a well-established in vitro model derived from a human neuroblastoma tumor. It is a subclone of the SK-N-SH cell line, commonly used for research on neuronal development, neurodegenerative diseases, and the effects of various compounds on neuronal cells.
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Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
2.5g l trypsin 1mmol l edta solution, by Nacalai Tesque (1 mentions)
Penicillin streptomycin solution, by Nacalai Tesque (1 mentions)
Fetal bovine serum (fbs), by Lonza (1 mentions)
Dulbecco s modified eagle s medium, by Merck Group (1 mentions)
High glucose and l glutamine, by Nacalai Tesque (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Accutase, by Nacalai Tesque (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
M7145, by Merck Group (1 mentions)
F 12 ham, by Merck Group (1 mentions)
Sh sy5y cells, by Thermo Fisher Scientific (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
19 protocols using sh sy5y neuroblastoma cell line
1
Glucose-6-phosphate dehydrogenase deficiency study
2
SH-SY5Y Neuroblastoma Cell Differentiation
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Dulbecco’s Modified Essential Medium/Ham’s Nutrient Mixture (DMEM:F12), Penicillin-Streptomycin solution and 2.5g/l-Trypsin/1mmol/l-EDTA Solution were purchased from Nacalai Tesque (Tokyo, Japan). Fetal bovine serum (FBS) and non-essential amino acids (NEAA) was purchased from Gibco-BRL (Grand Island, NY). Lipopolysaccharide (LPS) from Escherichia coli O55:B5 was purchased from Merck (Darmstadt, Germany).
SH-SY5Y neuroblastoma cell line were purchased from ATCC (ATCC® CRL-2266™). The cells were initially grown in Dulbecco’s Modified Essential Medium (DMEM:F12) which contains 4.5 g/l glucose with 2mM of L-glutamine and sodium pyruvate, supplemented with 15% FBS, 1% of Penicillin-Streptomycin mixed solution and 1% NEAA at 37°C with 5% carbon dioxide (CO2). Then, the cells were induced with 10 μM of all-trans retinoic acid for 5 days in a differentiation media (DMEM:F12, supplemented with 2.5% Fetal bovine serum (FBS) and 1% of Penicillin-Streptomycin mixed solution) (Forster et al., 2016 (link); Izham et al., 2018 (link)).
SH-SY5Y neuroblastoma cell line were purchased from ATCC (ATCC® CRL-2266™). The cells were initially grown in Dulbecco’s Modified Essential Medium (DMEM:F12) which contains 4.5 g/l glucose with 2mM of L-glutamine and sodium pyruvate, supplemented with 15% FBS, 1% of Penicillin-Streptomycin mixed solution and 1% NEAA at 37°C with 5% carbon dioxide (CO2). Then, the cells were induced with 10 μM of all-trans retinoic acid for 5 days in a differentiation media (DMEM:F12, supplemented with 2.5% Fetal bovine serum (FBS) and 1% of Penicillin-Streptomycin mixed solution) (Forster et al., 2016 (link); Izham et al., 2018 (link)).
3
SH-SY5Y Cells Response to MPP+ and PSELT
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The human SH-SY5Y neuroblastoma cell line (ATCC, Manassas, USA) was maintained in Dulbecco's modified Eagle's medium (Sigma-Aldrich, Saint-Quentin Fallavier, France), supplemented with 10% fetal bovine serum (Lonza, Levallois, France), 1% l -glutamine, 50 units/ml of penicillin and 50 units/ml of streptomycin (Thermo Fisher Scientific, Villebon-sur-Yvette, France), at 37 °C in 5% CO2 humidified atmosphere. The medium was renewed every 2–3 days. Twenty-four hours after plating, cells were treated or not with 500 μM or 1 mM MPP+ (Sigma-Aldrich) for 36 h in the presence or absence of PSELT (10 μM, dissolved in culture medium). The EZH2 inhibitor EPZ-6438 (Clinisciences, Nanterre, France), when present, was added at 10 μM at the same time as the peptide and MPP+. For the microarray gene expression analysis, cells were treated with PSELT for 6 h only.
4
SH-SY5Y Neuroblastoma Cell Cultivation
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The SH-SY5Y neuroblastoma cell line procured from ATCC (Manassas, VA, USA; Cat. #: CRL-2266) was cultivated in 25 cm2 cell culture flasks at 37 °C under a moistened atmosphere of 5% CO2 in Dulbecco’s modified Eagle medium (DMEM) with high glucose and L-glutamine (Nacalai Tesque, Kyoto, Japan; Cat. #: 08469-35), supplemented with 10% fetal bovine serum (FBS) (Thermo Fisher Scientific, Waltham, MA, USA; Cat. #: 10100147) and 1% penicillin-streptomycin (MilliporeSigma, Burlington, MA, USA; Cat. #: P0781). The media were replenished every 2–3 days, and at a confluence of 70–90%, cells were harvested with Accutase (Nacalai Tesque, Kyoto, Japan; Cat. #: 12679-54), sub-cultivated into cell culture vessels, and replated for immunocytochemical and cell death assays.
5
Culturing SH-SY5Y Neuroblastoma Cells
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SH‐SY5Y neuroblastoma cell line was obtained from ATCC (CRL‐2266). Cells were cultured in 1% pen/strep treated EMEM (Sigma, M4655) and F12 Ham (Sigma, N6658) mixed 1:1, 1% l ‐glutamine (200 mM; Gibco, 25030149), 1% Non‐essential amino acids (SAFC, M7145), and 10% FBS (Sigma, F4135). Before any assay, cells were thawed and plated in a T75 flask. During subculture, cell pellet was resuspended in 10 ml fresh medium and re‐plated. Cells were maintained in 5% CO2, 37°C. Cell passaging was done by incubating at 37°C with trypsin (Gibco, 12605028) for 5 min. trypsin was then inhibited 1:1 with fresh medium, and cells are then centrifuged. The pellet was washed with 5 ml PBS, resuspended in fresh medium, and plated.
6
SH-SY5Y Neuroblastoma Cell Culture
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SH-SY5Y neuroblastoma cell line was purchased from ATCC (Bethesda, MD, USA). SH-SY5Y cells were cultured in Dulbecco's modified Eagle's media (DMEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS), 1% L-glutamine and 100 U/ml penicillin/streptomycin (all from Gibco, Grand Island, NY, USA) and incubated in a humidified atmosphere of 95% under 5% of CO2 at 37°C. In differentiation studies, the medium was changed to DMEM containing 1% FBS, 1% L-glutamine and 100 U/ml penicillin/streptomycin (starvation culture medium) with or without metformin.
7
SH-SY5Y Neuroblastoma Cell Differentiation
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The SH-SY5Y neuroblastoma cell line was purchased from ATCC. Cells were grown in Dulbecco’s modified Eagle’s medium/nutrient mixture F-12 (DMEM/F12; Thermo Fisher Scientific, 11320–033), 2 mM L-glutamine (Thermo Fisher Scientific, 25030081), 100 U/mL penicillin, 100 μg/mL streptomycin (Thermo Fisher Scientific, 15140122), and 10% fetal bovine serum (FBS; GIBCO, 16140071). Cells were maintained at 37 °C in a saturated humidity atmosphere containing 95% air and 5% CO2. The cellular differentiation protocol has been previously described [43 (link), 64 (link)]. In brief, cells were seeded at an initial density of 104 cells/cm2 in culture dishes with complete medium. Trans retinoic acid (RA; Sigma-Aldrich, PHR1187) was added the day after plating at a final concentration of 10 µM in DMEM/F12 with 3% FBS, and cells were incubated for 7 days. The medium was replaced every other day, and on day 7, cells were harvested and used for experiments.
8
Culturing SH-SY5Y and Primary Entorhinal Neurons
9
SH-SY5Y Neuroblastoma Cell Culture
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The SH-SY5Y neuroblastoma cell line was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). The cell line was cultured as monolayers in a 1:1 ratio of ATCC-formulated Ham's F-12 Nutrient Mixture (F12) and Dulbecco's Modified Eagle Medium (DMEM, Sigma-Aldrich) containing 10 % heat-inactivated fetal bovine serum (FBS, GIBCO, Gaithersburg, MD, U.S.A.), essential amino acid, sodium pyruvate, 100 U/mL penicillin, and 100 μg/mL streptomycin at 37 °C in a humidified atmosphere containing 5 % CO2 in an incubator along the experiments. Cells were seeded at an initial density of 104 cells/cm2 in culture dishes (Corning, NY, U.S.A.). The medium was changed every 48 h. The cells were used at a low passage number (<35).
10
SH-SY5Y and 293T Cell Culture
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The human SH-SY5Y neuroblastoma cell line was purchased from the American Type Culture Collection (ATCC; cat. no. CRL-2266). The authenticity of the cells was confirmed using the Short Tandem Repeat assay report available at https://www.procell.com.cn/view/1401.html . The cell line was cultured according to the ATCC protocol. Briefly, cells were maintained in Eagle's Minimum Essential Medium (EMEM; cat. no. M4655; MilliporeSigma) supplemented with 10% FBS (cat. no. 12103C; MilliporeSigma) in a humidified incubator at 37°C with 5% CO2. The 293T cells were also purchased from the ATCC and maintained in DMEM (cat. no. D0819; MilliporeSigma) supplemented with 10% FBS in a humidified incubator at 37°C with 5% CO2. The 100 U/ml penicillin and 100 µg/ml streptomycin (cat. no. 10378016; Gibco; Thermo Fisher Scientific, Inc.) were added in EMEM and DMEM.
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