We used three human NB cell lines purchased from ATCC®: CRL22271 (SK-N-BE.2), CRL2266 (SH-SY5Y) and HTB11(SK-N-SH). CRL2266 and CRL2271 cells were cultured in 1:1 mixture of ATCC-formulated Eagle's Minimum Essential Medium (EMEM) and F12 Medium with 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 10 μg/ml streptomycin. HTB11 cells were cultured in Eagle's Minimum Essential Medium (EMEM) containing 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 10 μg/ml streptomycin. All cells were placed in a humidified atmosphere of 5% CO2 at 37°C and the medium was changed every 72 hours.
Crl 2266
Manufactured by American Type Culture Collection
Sourced in United States
The CRL-2266 is a cell line derived from human bone marrow cells. It is a myeloma plasma cell line that can be used for the production of monoclonal antibodies.
Automatically generated - may contain errors
Lab products found in correlation
Sh sy5y, by American Type Culture Collection (7 mentions)
Fetal bovine serum (fbs), by Merck Group (5 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Crl 2271, by American Type Culture Collection (3 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (3 mentions)
Crl 1573, by American Type Culture Collection (3 mentions)
Sh sy5y cell, by American Type Culture Collection (3 mentions)
Htb 11, by American Type Culture Collection (2 mentions)
Htb 22, by American Type Culture Collection (2 mentions)
Ccl 2, by American Type Culture Collection (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
L glutamine penicillin streptomycin solution, by Merck Group (2 mentions)
Ccl 127, by American Type Culture Collection (2 mentions)
Sh sy5y cell line, by American Type Culture Collection (2 mentions)
Fetal bovine serum (fbs), by American Type Culture Collection (2 mentions)
Complete emem medium, by American Type Culture Collection (2 mentions)
L15b300 medium, by Merck Group (2 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Dmem f12 medium, by Thermo Fisher Scientific (1 mentions)
37 protocols using crl 2266
1
Culturing Human Neuroblastoma Cell Lines
2
Cell Culture of Breast and Neuroblastoma Cells
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Human breast cancer cells (MCF-7, ATCC® HTB-22™) and human neuroblastoma cells (SH-SY5Y, ATCC® CRL-2266™) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) (Sigma-Aldrich, Dorset, UK) with 50 μM of glutamine, supplemented with 100 U/mL of penicillin/streptomycin (Sigma-Aldrich, Dorset, UK) and 100 mg/mL of 10% Fetal Bovine Serum (FBS) (Sigma-Aldrich, Dorset, UK). The cells were incubated in a humidified controlled atmosphere with a 95 to 5% ratio of air/CO2 at 37 °C.
3
Argon Preconditioning Protects Neuroblastoma Cells
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Neuroblastoma cells (cell line SH-SY5Y) were obtained directly from ATCC (Cat# CRL-2266, RRID: CVCL_019, Manassas, VA, USA), cultured in DMEM/F12 medium (GIBCO Life Technologies, Darmstadt, Germany) supplemented with 1% penicillin/streptomycin and 10% fetal calf serum and incubated in a humidified atmosphere containing 5% carbon dioxide at 37°C constant temperature until the cells reached 80% confluence. The cells for experiments were seeded in 6-well culture plates at a density of approximately 1.5 × 105 per well and cultured for a further 48 hours. Prior to rotenone treatment, the cells were exposed to gas mixtures containing argon at 25%, 50%, or 74% for 2 or 4 hours in a designated incubator with a humidified atmosphere. The gas delivery and mixing were done automatically, and the pre-set target values were reached in less than 60 seconds. The temperature was maintained at 37°C during all exposures. Subsequently, the cells were transferred to media containing 1% fetal calf serum to prevent the inactivation of rotenone by protein binding. Then, the rotenone treatment (20 µM) was performed. OxPAPC (30 µg/mL) was added 60 minutes before the argon treatment in the specific experiments. Cells were collected immediately after 4-hour rotenone treatment for FACS analysis and protein quantification.
4
Maintenance of Human Cancer and Stromal Cell Lines
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Human hepatocellular carcinoma HepG2 (ATCC® HB-8065™ ), human cervix adenocarcinoma HeLa (ATCC® CCL-2™ ), human gastric adenocarcinoma AGS (ATCC® CRL-1739™ ) and human colorectal adenocarcinoma HCT-15 (ATCC® CCL-225™) cell lines were maintained in RPMI-1640 culture media supplemented with 10% FBS (v/v), L-glutamine (2 mM), penicillin (50 U/mL) and streptomycin (50 µg/mL). Murine bone marrow stromal S17 cell line was kindly provided by D. Rawlings, UCLA, Los Angeles, CA. The latter cell line as well as human umbilical vein endothelial HUVEC (ATCC® CRL-1730™) and human neuroblastoma SH-SY5Y (ATCC® CRL-2266™ ) cell lines were grown in DMEM culture media supplemented with 10% FBS (v/v), L-glutamine (2 mM), penicillin (50 U/mL) and streptomycin (50 µg/mL). All cells were grown in an incubator at 37 °C and 5.0% CO2 in humidified atmosphere.
5
Paraquat-induced Neuroblastoma Cell Stress
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Human neuroblastoma (SH‐SY5Y) cells were grown at 37°C and 5% CO2 in a 1:1 mixture of EMEM and Ham's F12 medium (Gibco BRL) with 2 mM l ‐glutamine (Gibco BRL), 10% fetal bovine serum (FBS). Differentiation of these cells was carried out by adding 10 mM of retinoic acid (RA) twenty‐four hours after plating the cells. After 5 days in the presence of RA, cells were subjected to paraquat (PQ) exposure by incubation with different concentrations of PQ (Sigma) in complete media for 24 h at 37°C, with or without neuroprotective agents.
SH‐SY5Y cell line was purchased from ATCC in 2017 (CRL2266), HEK293 cells, passage 2 received from the Meshorer laboratory that purchased it from ATCC 2014 (CRL‐1573).
SH‐SY5Y cell line was purchased from ATCC in 2017 (CRL2266), HEK293 cells, passage 2 received from the Meshorer laboratory that purchased it from ATCC 2014 (CRL‐1573).
6
Characterization of Neuroblastoma Cell Lines
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Five NB cell lines were purchased from ATCC (New York, NY). CRL-2267, CCL-127, CRL-2271 (of male origin) CRL-2266, and CRL- 2149 (of female origin) were cultured in Eagle's Minimum Essential Media (EMEM) (ATCC, Manassas, VA) supplemented with 10% heat inactivated Fetal Bovine Serum (FBS), and l -Glutamine-penicillin-streptomycin solution (designated complete media) (Sigma, St. Louis, MO) at 37 °C, 5% CO2. SMS-KCNR (of male origin) was purchased from Children's Oncology Group (Texas Tech University) and was cultured in RPMI-1640 (Sigma). Two terminally differentiated olfactory neuroblastoma cell lines, T-268 and JFEN, were a kind gift from Timothy J. Triche (Department of Pathology, Children's Hospital of Los Angeles, Los Angeles, CA 90027; 21). The stock cell cultures were grown in 25 mL or 75 mL flasks (Thermo-Scientific, Nunc, Rochester, NY). ZIKV strains PRVABC59 and MR 766 were provided by the CDC (kind gifts from Brandy Russel, Fort Collins, CO) and flavivirus envelop antibody (FE1), which cross-reacts with ZIKV, was purchased from Invitrogen (Cat. # MA1-71258). Two NB cell lines CRL-2266 and CRL-2267 were exposed to retinoic acid (RA) at 1 μM final concentration for 48 h (Breitman et al., 1980 (link)). Both cell lines exhibited partial differentiation as measured by morphologically and by immunostaining with anti-tubulin antibody.
7
Culturing Human Neuroblastoma SH-SY5Y Cells
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The human neuroblastoma SH-SY5Y cell line was obtained from the American Type Culture Collection (CRL-2266™, ATCC, Manassas, VA, USA). The cells were cultured in a nutrient mixture consisting of Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, Waltham, MA, USA) supplemented with 2 mM L-glutamine, 100 U/mL penicillin–streptomycin, and 10% heat-inactivated fetal bovine serum (FBS), which were all obtained from Gibco (Gibco, Waltham, MA, USA). The culture was maintained in a humidified atmosphere at 37 °C with 5% CO2 according to the protocol outlined in the literature [24 (link)].
8
Culture of Human Dermal Fibroblasts and Neuroblastoma Cells
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The human dermal fibroblast cell (HDFa, ATCC® PCS-201-012™) culture was grown in the DMEM medium supplemented with 1% penicillin/streptomycin, 10% FBS and 5% CO2 at a 37 °C temperature until the cultures reached 80% confluency. The human neuroblastoma cell (SH-SY5Y, ATCC® CRL-2266™) line was cultured in DMEM/F12 culture media containing 1% penicillin/streptomycin and 10% fetal bovine serum at 37 °C and 5% CO2. Neuroblastoma cell cultures were incubated to grow up to 80% cell density.
9
Hypoxic Culture of SH-SY5Y Cells
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Human neuron-like SH-SY5Y neuroblastoma cells (ATCC® CRL-2266™) were placed in the RPMI-1640 medium containing 2 mM L-glutamine supplemented with 10% heat-inactivated fetal bovine serum and 100 U/ml penicillin/streptomycin. The culture was kept in a standard wet incubator with 5% CO2 at 37°C, and the original medium was replaced with fresh medium once every 2 days. When 90% cells were fused, the medium was divided as 1:4. Cells were placed in the calibration gas containing 1% O2 or 3% O2 (the concentration of CO2 was adjusted to 5% under these two conditions) and the cells were placed in a humidified microaerophilic culture system (DWS HypOxystation) to prepare the anaerobic environment. The cells were kept in an incubator at 37°C at different times. Control culture was kept for the same time under normal oxygen content.
10
Hypoxia-Reoxygenation Cell Culture
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Human monocytic cell line THP-1 cells obtained from ATCC (1 × 106 cells/mL) were suspended in a culture dish containing RPMI 1640 medium. Human umbilical vein endothelial cells (HUVECs) purchased from Lonza (Basel, Switzerland) were cultured in Clonetics Endothelial Cell Growth Media supplemented with the BulletKit containing bovine brain extract, epidermal growth factor, hydrocortisone, gentamicin, amphotericin B, 2% fetal bovine serum, 1% penicillin/streptomycin and ascorbic acid. Human SH-SY5Y neuron cells purchased from ATCC (® CRL-2266™) were cultured in ATCC-formulated MEM/F12 (1:1) growth medium supplemented with 10% fetal bovine serum, 1x non-essential amino acids, 1 mM sodium pyruvate, 1.5 g/L sodium bicarbonate, and 1% penicillin/ streptomycin. Cells were exposed to IHR or normoxic conditions in custom-designed, incubation chambers which were attached to an external O2–CO2 hand-driven controller. IHR protocol consists of a 25-min hypoxic period (0% O2 and 5% CO2) and 35 min of the re-oxygenation period (21% O2 and 5% CO2) per cycle, 8 cycles /day for 3 days, which has been shown to achieve an episodic decrease in culture medium SaO2 by 30–40% [29 (link)].
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