Sk n mc cells

Manufactured by American Type Culture Collection

Sourced in United States

SK-N-MC cells are a human-derived cell line originating from a neuroblastoma. They are adherent cells that can be used for various cell biology and molecular biology research applications.

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13 protocols using sk n mc cells

Synthesis and Characterization of αCGRP Analogues

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Human αCGRP and CGRP analogues (Bachem AG, Switzerland) were dissolved in pure water and kept frozen (−18 °C) in aliquots prior to use.
All Fmoc L- and D-amino acids (CEM), Rink Amide ProTide resin (CEM), diisopropylcarbodiimide (DIC; Apollo Scientific), Oxyma Pure™ (CEM), N.N’-dimethylformamide (DMF; Fisher Scientific), diisopropylethylamine (DIPEA; Merck Millipore) and piperidine (Merck Millipore) were purchased from commercial suppliers and used directly as indicated in the appropriate experimental procedures. SK-N-MC cells were purchased from ATCC. Earlés Balanced Salts, L-glutamine, fetal bovine serum (FBS), sodium pyruvate and non-essential amino acids were purchased from Life Technologies (Sweden). HitHunter cyclic AMP assay kits were purchased from DiscoveRx (Eurofins). All other reagents (trifluoroacetic acid (TFA), triisopropylsilane (TIPS), chitosan [CAS number 9012–76-4] were purchased from Sigma Aldrich and solvents (HPLC grade) were purchased from Fisher Scientific.

von Mentzer B., Russo A.F., Zhang Z., Kuburas A., Killoran P.M., D’Aloisio V., Nizic L., Capel V., Kendall D.A., Coxon C.R, & Hutcheon G.A. (2020). A CGRP receptor antagonist peptide formulated for nasal administration to treat migraine. The Journal of Pharmacy and Pharmacology, 72(10), 1352-1360.

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RET-expressing SK-N-MC Cell Line Protocol

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SK-N-MC cells stably expressing either a RET expression construct containing the extracellular and transmembrane domains of the EGF receptor linked to either the RET9 isoform, RET51 isoform intracellular domains or an empty vector were kindly provided by Dr. Michael Skinner (UT Southwestern Medical Center, Dallas, TX). Unmodified SK-N-MC cells are an established cell line obtainable from ATCC (Manassas, VA). Cells were maintained at 95% air and 5% CO₂ at 37°C and grown in MEMS medium (Life Technologies, Grand Island, NY) supplemented with 10% FBS (Atlanta Biologicals, Flowery Branch, GA), pen/strep, and 400 μg of G418 (Hyclone, Logan, UT) for continued selection. Cells were routinely checked for RET expression by western blot analysis. TT cells (CLR-1803) were obtained from ATCC and maintained in F-12K medium with 10% FBS and pen/strep.

Burdine L.J., Burdine M.S., Moreland L., Fogel B., Orr L.M., James J., Turnage R.H, & Tackett A.J. (2015). Proteomic Identification of DNA-PK Involvement within the RET Signaling Pathway. PLoS ONE, 10(6), e0127943.

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Culturing Diverse Human Cell Lines

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The HeLa cell line was obtained from the American Type Culture Collection (ATCC CCL-2; Manassas, VA, USA) and was maintained in minimum essential medium (MEM; Corning Cellgro; Manassas, VA, USA) supplemented with 10% fetal bovine serum (FBS; Sigma Aldrich; St. Louis, MO, USA) and 1% penicillin–streptomycin solution (Hyclone; Logan, UT, USA). SK-N-MC cells were obtained from ATCC (HTB-10) and maintained in MEM supplemented with 10% FBS (Sigma Aldrich; St. Louis, MO, USA), 1% penicillin–streptomycin solution (Hyclone; Logan, UT, USA), 1% sodium pyruvate (Hyclone; Logan, UT, USA), and 1% non-essential amino acid solution (Hyclone; Logan, UT, USA). HUH-7 cells were obtained from the JCRB Cell Bank (JCRB0403) and were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM; Corning Cellgro; Manassas, VA, USA) supplemented with 5% FBS (Sigma Aldrich; St. Louis, MO, USA) and 1% penicillin–streptomycin solution (Hyclone; Logan, UT, USA). Finally, Vero cells (ATCC CCL-81) were maintained in MEM (Corning Cellgro; Manassas, VA, USA) supplemented with 5% FBS (Sigma Aldrich; St. Louis, MO, USA) and 1% penicillin–streptomycin solution (Hyclone; Logan, UT, USA). Cells were maintained at 37 °C and 5% CO₂. Cells were split twice weekly to maintain subconfluency.

Franco E.J., Cella E., Tao X., Hanrahan K.C., Azarian T, & Brown A.N. (2023). Favipiravir Suppresses Zika Virus (ZIKV) through Activity as a Mutagen. Microorganisms, 11(5), 1342.

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Cell Culture Protocol for SK-N-MC and N2A

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SK‐N‐MC cells (ATCC) and N2A cells (ATCC) were cultured according to the vendor's protocol. In brief, cells were grown at 37C in 5% CO2. The used medium for both cell lines is ATCC‐formulated Eagle's Minimum Essential Medium (Catalog No. 30–2003). 1% Pen/Strep and foetal bovine serum to a final concentration of 10% was added.

Russo T., Kolisnyk B., B. S. A., Plessis‐Belair J., Kim T.W., Martin J., Ni J., Pearson J.A., Park E.J., Sher R.B., Studer L, & Riessland M. (2024). The SATB1‐MIR22‐GBA axis mediates glucocerebroside accumulation inducing a cellular senescence‐like phenotype in dopaminergic neurons. Aging Cell, 23(4), e14077.

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Culturing SK-N-MC and Neuro-2A Cells

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SK-N-MC cells (ATCC) and Neuro-2A (N2A) cells (ATCC) were cultured according to the vendor’s protocol. In brief, cells were grown at 37C in 5% CO2. The used medium for both cell lines is ATCC-formulated Eagle’s Minimum Essential Medium (Catalog No. 30-2003). 1% Pen/Strep and fetal bovine serum to a final concentration of 10% was added.

Russo T., Kolisnyk B., Aswathy B., Wan Kim T., Martin J., Plessis-Belair J., Ni J., Pearson J.A., Park E.J., Sher R.B., Studer L, & Riessland M. (2023). The SATB1-MIR22-GBA axis mediates glucocerebroside accumulation inducing a cellular senescence-like phenotype in dopaminergic neurons. bioRxiv.

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Synthesis and Characterization of αCGRP Analogues

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Human αCGRP and CGRP analogues (Bachem AG, Budendorf, Switzerland) were dissolved in pure water and kept frozen (−18 °C) in aliquots before use.
All Fmoc l- and d-amino acids (CEM Microwave Technology Ltd, Buckingham, UK), Rink Amide ProTide resin (CEM), diisopropylcarbodiimide (DIC; Apollo Scientific, Stockport, UK), Oxyma Pure (CEM), N.N′-dimethylformamide (DMF; Fisher Scientific, Loughborough, UK), diisopropylethylamine (DIPEA; Merck Millipore, Watford, UK) and piperidine (Merck Millipore) were purchased from commercial suppliers and used directly as indicated in the appropriate experimental procedures. SK-N-MC cells were purchased from ATCC (Manassas, VA, USA). Earle’s Balanced Salts, l-glutamine, fetal bovine serum (FBS), sodium pyruvate and non-essential amino acids were purchased from Life Technologies (Stockholm, Sweden). HitHunter cyclic AMP assay kits were purchased from DiscoveRx (Eurofins, Fremont, CA, USA). All other reagents (trifluoroacetic acid (TFA), triisopropylsilane (TIPS), chitosan [CAS number 9012-76-4]) were purchased from Sigma-Aldrich (Gillingham, UK), and solvents (HPLC grade) were purchased from Fisher Scientific (Loughborough, UK).

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Culturing and Differentiating Human NPC Cells

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Cell culture and media The human NPC cell line hNS-1 was generously donated from Centro de Biología Molecular BSevero Ochoa,^Madrid, Spain. These cells were grown in poly-L-lysine (Sigma-Aldrich, St. Louis, MO)-coated plastic flasks using a cell culture medium that contained DMEM-F12 (1:1, Gibco, Invitrogen Life Technologies, Carlsbad, CA) supplemented with 0.6 % glucose, N2 (Life Technologies), 0.5 % AlbuMAX I (Gibco), and human epidermal and fibroblast growth factors (EGF/FGF-2, R&D Systems) as previously described (Villa et al. 2000) . For differentiation experiments, hNS-1 cells were cultured 5 days without FGF-2 and EFG and in the presence of fetal bovine serum 0.5 % (FBS, ATCC, Rockville, MD). SK-N-MC cells (ATCC, Lot 4399800, Manassas, VA) were grown in MEM (ATCC) supplemented with 10 % FBS. Human foreskin fibroblasts were obtained from primary culture and were grown in DMEM medium supplemented with 10 % calf serum (Gibco). MDCK cells (ATCC) were grown in the same conditions as fibroblasts. Penicillin and streptomycin were used (100 U/mL and 100 μg/mL) to prevent culture contamination. For cells requiring serum, a maintenance medium was used where the serum concentration was decreased to 2 % after CMV inoculation. All cell cultures were grown at 37 °C in a 95 % humidity and a 5 % CO 2 atmosphere. Cell karyotyping and mycoplasma contamination were assessed periodically.

González-Sánchez H.M., Monsiváis-Urenda A., Salazar-Aldrete C.A., Hernández-Salinas A., Noyola D.E., Jiménez-Capdeville M.E., Martínez-Serrano A, & Castillo C.G. (2015). Effects of cytomegalovirus infection in human neural precursor cells depend on their differentiation state. Journal of neurovirology, 21(4).

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Culturing SK-N-MC Cells with Transfection

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SK-N-MC cells (American Type Culture Collection) were cultured in the Dulbecco's Modified Eagle Medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin in a 37°C incubator with 5% CO₂. All plasmid transfections were carried out by using Lipofectamine 2000 (Thermo Fisher Scientific). All siRNAs were transfected with Lipofectamine^® RNAIMAX (Thermo Fisher Scientific) transfection reagent.

An Y., Chen Z.S., Chan H.Y, & Ngo J.C. (2022). Molecular insights into the interaction of CAG trinucleotide RNA repeats with nucleolin and its implication in polyglutamine diseases. Nucleic Acids Research, 50(13), 7655-7668.

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Transfection and Peptide Application in SK-N-MC Cells

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SK-N-MC cells (American Type Culture Collection [ATCC]) were cultured in DMEM (GE Healthcare BioSciences) supplemented with 10% fetal bovine serum and 1% penicillin–streptomycin. The cells were maintained in a 37°C humidified cell culture incubator supplemented with 5% CO₂. DNA constructs and siRNA were transfected with Lipofectamine 2000 (Thermo Fisher Scientific) and Lipofectamine RNAiMAX (Thermo Fisher Scientific), respectively. After transfection, TAT-BIND or TAT-BIND-FITC peptides were applied with Opti-MEM medium (Thermo Fisher Scientific).

Peng S.I., Leong L.I., Sun J.K., Chen Z.S., Chow H.M, & Chan H.Y. (2022). A peptide inhibitor that rescues polyglutamine-induced synaptic defects and cell death through suppressing RNA and protein toxicities. Molecular Therapy. Nucleic Acids, 29, 102-115.

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Neuroblastoma Cell Viability Assay

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SK-N-MC cells (human neuroblastoma cell line) were obtained from the American Type Culture Collection (Bethesda, MD, USA). Cells were cultured in Minimal Eagle’s Medium (MEM; Gibco) supplemented with 10% fetal calf serum, antibiotics (100 units/mL penicillin, 100 µg/mL streptomycin), 2-mM l-glutamine and kept in a humidified air containing 5% CO₂ at 37 °C. Transient transfections were carried out using Lipofectamine 3000 reagent (Thermo Fisher Scientific, Waltham, MA, USA), according to manufacturer’s instruction. For viability assays, cells were treated with tetrazolium salt methyl-thiazol-tetrazolium (MTT) for 30 min, and then analyzed spectrophotometrically at 550 nm. Viability was determined as percent of control cells treated with vehicle alone.

Chang C.C., Li H.H., Tsou S.H., Hung H.C., Liu G.Y., Korolenko T.A., Lai T.J., Ho Y.J, & Lin C.L. (2020). The Pluripotency Factor Nanog Protects against Neuronal Amyloid β-Induced Toxicity and Oxidative Stress through Insulin Sensitivity Restoration. Cells, 9(6), 1339.

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