The LAN-5 neuroblastoma cell line was kindly provided by Dr Stuart Elliott Siegel (Children’s Hospital Los Angeles, Los Angeles, CA, USA) and the origin has been described previously (17 (link),18 (link)); the SK-N-SH cell line was purchased from American Type Culture Collection (Cambridge, MA, USA). The cells were maintained in RPMI-1640 (Gibco, Paisley, UK) supplemented with 10% fetal calf serum (FCS; Gibco) in an atmosphere of 5% CO2 at 37°C.
Sk n sh cell line
Manufactured by American Type Culture Collection
Sourced in United States
The SK-N-SH cell line is a neuroblastoma cell line derived from a human female patient. It is commonly used for research purposes in the study of neuroblastoma and related neurological conditions.
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Lab products found in correlation
Penicillin streptomycin neomycin, by Merck Group (2 mentions)
Sk n sh, by Merck Group (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Sh sy5y cell, by American Type Culture Collection (1 mentions)
N acetylcysteine, by Merck Group (1 mentions)
Tert butyl hydroperoxide tbhp, by Merck Group (1 mentions)
Fluo 3 am, by Dojindo Laboratories (1 mentions)
Lactate dehydrogenase ldh cytotoxicity assay kit, by Beyotime (1 mentions)
T75 flask, by Corning (1 mentions)
Retinoic acid, by Merck Group (1 mentions)
1 methyl 4 phenylpyridinium ion mpp, by Merck Group (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Sk n sh, by Thermo Fisher Scientific (1 mentions)
7 protocols using sk n sh cell line
1
Cell Line Maintenance Protocol
2
Differentiation of SK-N-SH cells into neuronal cells
3
Polystyrene Nanoparticle Toxicity in Neuroblastoma Cells
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The 50 nm polystyrene nanoparticle (PS-NPs) beads (5% w/v) were purchased commercially from Janus New-Materials (Nanjing, China) (refer to the Supplementary Materials for more detailed parameters). SHSY-5Y cells, the clonal subline of neuroblastoma SK-N-SH cell line, were obtained from American Type Culture Collection (ATCC, Rockville, MD, USA). The 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di- phenytetrazo-liumromide (MTT), Rhodamine 123 (Rh 123), 2,7-dichlorodi- hydrofluorescein diacetate (DCFH-DA), tert-Butyl hydroperoxide (tBHP), and N-Acetylcysteine (NAC) were obtained from Sigma (St. Louis, MO, USA). The annexin V-fluoresceine isothiocyanate/propidium iodide (annexin V-FITC/PI) apoptosis detection kit was obtained from Sungene (Tianjin, China). Protein extraction and assay kit was purchased from Thermo (MA, USA). Fluo 3-AM was acquired from Dojindo (Kyushu, Japan). Lactate dehydrogenase (LDH) cytotoxicity assay kit and adenosine 5′-triphosphate (ATP) detection kit were obtained from Beyotime (Shanghai, China).
4
Differentiating SK-N-SH Cells into Neuron-Like Cells
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Human SK-N-SH cell line was obtained from American Type Culture Collection (ATCC #HTB-11). Cells were cultured in Eagle’s Minimum Essential Medium (EMEM; ATCC). This media was supplemented with 1 % penicillin-streptomycin-neomycin (Sigma) and 10 % (v/v) fetal bovine serum (FBS). Retinoic acid (RA; Sigma) was used to induce SK-N-SH cells [12 (link)] to differentiate into more neuron-like cells [13 (link)] because this cell line is a mixture of different cell types. Approximately 15,000 cells were cultured in T-75 flask (Corning) supplemented with EMEM media for two days, and then Retinoic acid (10 μM) was added. Cells were treated with RA for two weeks and media was replaced every three-four days [12 (link)]. Cultures were monitored visually using light microscopy for morphological changes, and evaluated for neuronal cell markers (NeuN, PSD95 and NCAM) during the differentiation process [12 (link), 14 (link)].
5
Neuroprotection Assay in SK-N-SH Cells
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Human neuroblastoma SK-N-SH cell line procured from American Type Culture Collection (ATCC, Manassas, VA, USA) was maintained in Eagle's Minimum Essential Medium (EMEM; ATCC, Manassas, VA, USA) decorated by 10% fetal bovine serum (FBS; Sijiqing Biologic Co. Ltd., Hangzhou, China) under the atmosphere of 5% CO 2 at 37°C. SK-N-SH cells were passaged at a 1:3 ratio every 3-4 days. SK-N-SH cells were cotreated by ascending concentrations of ICS II (12.5, 25 and 50 μM with 24 h; Nantong Feiyu Biological Technology Co., Ltd., Nanjing, China) [ 27 ] and 2 mM 1-methyl-4-phenylpyridinium ion (MPP + ) (Sigma-Aldrich, St. Louis, MO, USA) for 24 h [ 28 ] with the absence or presence of 10 μM Nrf2 inhibitor ML385 (Aladdin, Shanghai, China) for 24 h. [ 29 ]
6
Propagation of Neuronal Cell Lines
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SK-N-SH cell lines were obtained from the American Type Culture Collection (ATCC; Rockville, MD, USA) and the cell culture method has been previously described (Moon and Park, 2015 (link)). The mouse neuronal cell lines ZW 13–2 and Zpl 3–4, which were established from the hippocampus of ICR (Prnp+/+) and Zürich I Prnp−/− mice, respectively, were kindly provided by Professor Yong-Sun Kim (Hallym University, Chuncheon, Kangwon-do, South Korea). The cells were grown in Dulbecco’s Modified Eagle Medium (DMEM; HyClone, Logan, UT, USA) containing 10% fetal bovine serum (FBS) and gentamycin (0.1 mg/mL) in a humidified incubator maintained at 37 °C with 5% CO2.
7
Neuroblastoma SK-N-SH Cell Culture
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Human neuroblastoma (SK-N-SH) cell lines were purchased from the American Type Culture Collection (Manassas, VA, USA) and used to assess neuronal cell properties. SK-N-SH cells were grown in minimum essential medium (MEM) supplemented with 10 % FBS, 100 U/ml penicillin, and 100 µg/ml streptomycin (GIBCO-BRL, Gaithersburg, MD) at 37 °C in a humidified 5 % CO2 and 95 % air incubator. The cells were cultured to 80 % confluence and passaged every 3 days by trypsinization with 0.025 % trypsin/EDTA.
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