African green monkey kidney cells (Vero) and SH-SY5Y were obtained from the American Tissue-Culture Collection (ATCC) (Rockville, MD, USA). Veros were maintained on Dulbecco’s Modified Eagle Medium (Gibco-BRL, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Gibco-BRL, Grand Island, NY, USA) and 100 ug/mL Primocin© (Invitrogen, INC., Carlsbad, CA, USA). SH-SY5Y were maintained on Eagle’s Minimum Essential Medium (ATCC, Rockville, MD, USA) supplemented with 15% fetal bovine serum (Gibco-BRL, Grand Island, NY, USA) and 100 ug/mL Primocin© (Invitrogen, INC., Carlsbad, CA, USA). The wildtype (WT) and Δ481N viruses were a gift from Prashant Desai (Johns Hopkins University, Baltimore, MD, USA). The mutants were made on a KOS background with a GFP on UL37. Viruses were grown and titrated on Vero cells [23 (link)].
Sh sy5y
Manufactured by American Type Culture Collection
Sourced in United States, United Kingdom, Germany, China, France, Italy
The SH-SY5Y is a human cell line derived from a bone marrow biopsy. It is a subclone of the parental cell line SK-N-SH, which was originally isolated from a metastatic neuroblastoma tumor. The SH-SY5Y cell line is commonly used in neurobiological and neurodegenerative research.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (131 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (85 mentions)
Sk n sh, by American Type Culture Collection (64 mentions)
Sk n as, by American Type Culture Collection (60 mentions)
Penicillin, by Thermo Fisher Scientific (57 mentions)
Imr 32, by American Type Culture Collection (56 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (56 mentions)
Streptomycin, by Thermo Fisher Scientific (55 mentions)
Sh sy5y, by Thermo Fisher Scientific (53 mentions)
Hek293t, by American Type Culture Collection (42 mentions)
Hek293, by American Type Culture Collection (30 mentions)
Fetal bovine serum (fbs), by Merck Group (29 mentions)
Be 2 c, by American Type Culture Collection (25 mentions)
Sk n dz, by American Type Culture Collection (24 mentions)
Sk n be 2, by American Type Culture Collection (24 mentions)
Mcf 7, by American Type Culture Collection (24 mentions)
Dmem f12, by Thermo Fisher Scientific (23 mentions)
Sh sy5y, by Merck Group (21 mentions)
Glutamax, by Thermo Fisher Scientific (21 mentions)
Mda mb 231, by American Type Culture Collection (20 mentions)
655 protocols using sh sy5y
1
Vero and SH-SY5Y Cell Culture Protocol
2
Characterization of Human Cell Lines
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The human fetal astrocyte cell line SVGA, the human microglial cell line HMC3, and the human neuroblastoma cell line SH-SY5Y were used for our investigations. The SVGA cells were kindly provided by the group of Christine Hanssen Rinaldo, University Hospital of North Norway (Henriksen et al., 2014 (link)) with the permission of Altwood (Schweighardt et al., 2001 (link)). The HMC3 and SH-SY5Y cells were purchased from the American Type Culture Collection (ATCC, Manassas, Virginia, USA). SVGA and HMC3 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Life Technologies, Carlsbad, CA, USA). SH-SY5Y cells were cultured in a half and half mixture of F12 (ATCC) and Eagle’s minimum essential medium (EMEM). The corresponding media were supplemented with 10% fetal bovine serum (FBS; PAN-Biotech GmbH, Aidenbach, Germany), 1% penicillin–streptomycin (10,000 U/ml; Thermo Fisher Scientific, Waltham, MA, USA), and 2 mM of additional l -glutamine (Thermo Fisher Scientific). Purity of the cells was ascertained by immunostaining with cell type-specific markers and by the absence mycoplasma contamination. Cell line identity was verified by short tandem repeat profiling as previously described (Adamski et al., 2017 (link)).
3
Propagation and Infection of Cell Lines
4
Murine and Human Neuroblastoma Cell Culture
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The murine neuroblastoma cell line Neuro2a (N2a) is derived from an A/J mouse and was purchased from the American Type Culture Collection (ATCC, Manassas, Virginia, US). The aggressive N2a subclone AgN2a was derived from repeated in vivo passaging of these cells as described previously [14 (link)]. The cells were maintained in Dulbecco’s Modified Essential Medium (DMEM) supplemented with 1% penicillin-streptomycin (Invitrogen, Carlsbad, California, US) and 10% fetal bovine serum (Gemini Bioproducts, Sacramento, California, US). Mouse splenocytes were cultured in RPMI medium supplemented with 2 mM L-glutamine, 10% fetal bovine serum, and 1% penicillin-streptomycin. Cells were grown at 37°C under 5% CO2.
Human neuroblastoma cell lines IMR-32, SK-N-SH, and SH-SY5Y were obtained from the ATCC. IMR-32 and SK-N-SH cells were grown in ATTC Eagle’s Minimum Essential Medium (EMEM) supplemented with 10% FBS. SH-SY5Y cells were cultured in ATCC EMEM mixed 1:1 with F12 medium, and FBS was added to a final concentration of 10%.
Human neuroblastoma cell lines IMR-32, SK-N-SH, and SH-SY5Y were obtained from the ATCC. IMR-32 and SK-N-SH cells were grown in ATTC Eagle’s Minimum Essential Medium (EMEM) supplemented with 10% FBS. SH-SY5Y cells were cultured in ATCC EMEM mixed 1:1 with F12 medium, and FBS was added to a final concentration of 10%.
5
Differentiation of Human Neuroblastoma SH-SY5Y
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Human neuroblastoma SH-SY5Y (CLS Cat# 300154/p822_SH-SY5Y, RRID:CVCL_0019) was obtained from ATCC (Manassas, VA, USA). Cells with a neuronal-like phenotype were cultured as described in [19 (link)]. In brief, SH-SY5Y cells were grown in Basic Growth Media (DMEM/F12 (1:1) (ThermoFisher Scientific, Waltham, MA, USA) with 2 mM L-glutamine, 10% fetal bovine serum (HyClone, Logan, UT, USA), 100 U/mL penicillin, 100 µg/mL streptomycin at 37 °C/5% CO2. For cell differentiation, on day 0, cells were seeded in Basic Growth Media to the confluence of 40–50%; on day 1, the medium was changed to the differentiation medium (DMEM/F12 (1:1)) with 2 mM L-glutamine, 1% fetal bovine serum, 100 U/mL penicillin, 100 µg/mL streptomycin, 10 µM all-trans-retinoic acid (Sigma-Aldrich, St. Louis, MO, USA), and cells were protected from light. Patch–clamp experiments were performed on days 6–8.
6
Characterization of Cell Lines in Cancer Research
7
Cell Culture and Mouse Maintenance
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HEK293 (ATCC, CRL-3216), HeLa (ATCC, CCL-2), and SH-SY5Y (ATCC, CRL-2266) cells were purchased from ATCC. HEK293 and HeLa were maintained in Dulbecco’s modified Eagle’s medium (DMEM, Gibco, 11965118) + 10% fetal bovine serum (FBS, Gibco, 10082147) + penicillin–streptomycin (Gibco, 15140163), in 37 °C incubator with 5% CO2. SH-SY5Y was maintained in 1:1 mixture of Eagle’s Minimum Essential Medium and F12 Medium (ATCC, 30-2003) + 10% FBS + penicillin–streptomycin. Wild-type C57BL/6J mice were bred and maintained under specific pathogen-free conditions, fed standard laboratory chow, and kept on a 12-h light/dark cycle and temperature and humidity were kept at 22 ± 1 °C, 55 ± 5%. C57BL/6J female or male mice aged 4–6 weeks old were used for all experiments. All cell culture were handled according to protocols approved by the University of Southern California. All animals were used according to animal use protocols granted by the Institutional Animal Care and Use Committee at the University of Southern California.
8
Neuroblastoma Cell Lines and Xanthohumol Treatment
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Three human neuroblastoma (NB) cell lines (SK-N-AS, NGP, and SH-SY-5Y) were used in this study, and SK-N-AS and SH-SY-5Y were purchased from ATCC. The NGP was a kind gift from Dr. Thiele. The cells were grown in Roswell Park Memorial Institute Media (RPMI; Life Technologies, Carlsbad, CA), supplemented with 10% fetal bovine serum (Sigma-Aldrich, St. Louis, MO), 100 IU/mL penicillin, and 100 μg/mL streptomycin (Life Technologies) in a humidified atmosphere of 5% CO2 in air at 37°C. Xanthohumol (Tocris, Minneapolis, MN or Selleckchem, Houston, TX) was dissolved in dimethyl sulfoxide (DMSO: Sigma-Aldrich) to prepare stock solutions of 50 mM. An equivalent volume of DMSO alone served as a negative control. TRAIL/Apo-2L was purchased from PeproTech (Rocky Hill, New Jersey, USA) and stored in aliquots at -80°C.
9
Cell Culture Conditions for Multiple Cell Lines
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The following cell lines were acquired from ATCC: CHO-K1 (CRL-9618), Neuro-2a (CCL-131), SH-SY5Y (CRL-2266), and AR42J (CRL-1492). GT1-7 cells were a kind gift from the lab of Pamela Mellon (University of California, San Diego), while CHO cells stably expressing OX1 were generously provided by the lab of Patricia McDonald (The Scripps Research Institute). All cells were grown in the presence of 10% HI-FBS (Gibco) at 37°C, 5.0% CO2. Base media for each cell line are as follows: DMEM, high glucose (Gibco) for GT1-7 and CHO cells, Eagle's Minimum Essential Medium (ATCC) for Neuro-2a (N2A) and SH-SY5Y cells, and F-12K Medium (ATCC) for AR42J cells.
10
Cultivation and Differentiation of Human Cell Lines
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The human neuroblastoma cell line SHSY5Y (CRL-2266), the human astrocytoma cell line CCF-STTG1 (CRL-1718), human monocytic leukemia cell line THP-1 (TIB-202), and the human lung carcinoma cell line A549 (CCL-185) were obtained from the American Type Culture Collection (ATCC). SHSY5Y cells were cultured in a 1:1 mixture of EMEM (ATCC 30–2003) and F-12K (ATCC 30–2004) supplemented with 10% fetal bovine serum (FBS) (ATCC 30–2020). CCF-STTG1 and THP-1 cells were cultured in RPMI-1640 (ATCC 30–2001) supplemented with 10% FBS. A549 cells were cultured in F-12K (ATCC 30–2004) supplemented with 10% FBS. Cells were maintained in culture following the manufacturer’s recommendations. HIV-1 infected U937 Cells (U1; NIH AIDS Reagent Program) were cultured in RPMI-1640 supplemented with 10% exosome-depleted FBS. To generate MDMs, THP-1 cells and U1 cells were treated with phorbol 12-myristate 13-acetate (PMA; 100 nM) and incubated for four to five days. U1-MDMs then received two treatments of cART (emtricitabine, tenofovir disoproxil fumarate, ritonavir, darunavir; 2.5 µM each) over a five day incubation period.
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