Neuro 2a cells

Manufactured by American Type Culture Collection

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Neuro-2a cells are a mouse neuroblastoma cell line derived from a spontaneous tumor. They are commonly used as a model for studying neurological processes and function.

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Neuro-2a (124 citations)

75 protocols using neuro 2a cells

Mitochondrial Toxicity Assessment in Neuro2A Cells

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Example 6

Mitochondrial Toxicity Assays in Neuro2A Cells

To estimate the potential of the compounds of this invention to cause neuronal toxicity, mouse Neuro2A Cells (American Type Culture Collection 131) were used as a model system (see Ray A S, Hernandez-Santiago B I, Mathew J S, Murakami E, Bozeman C, Xie M Y, Dutschman G E, Gullen E, Yang Z, Hurwitz S, Cheng Y C, Chu C K, McClure H, Schinazi R F, Anderson K S. Mechanism of anti-human immunodeficiency virus activity of beta-D-6-cyclopropylamino-2′,3′-didehydro-2′,3′-dideoxyguanosine. Antimicrob. Agents Chemother. 2005, 49, 1994-2001). The concentrations necessary to inhibit cell growth by 50% (CC₅₀) were measured using the 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide dye-based assay, as described. Perturbations in cellular lactic acid and mitochondrial DNA levels at defined concentrations of drug were carried out as described above. In all experiments, ddC and AZT were used as control nucleoside analogs.

US09932326B2. Potent and selective inhibitors of hepatitis C virus (2018-04-03). Cocrystal Pharma, LLC [US], Emory University [US]. Inventors: Steven J. Coats [US], Franck Amblard [US], Hongwang Zhang [US], Longhu Zhou [US], Richard Anthony Whitaker [US], Tamara Rosario McBrayer [US], Raymond F. Schinazi [US], Junxing Shi [US].

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Mitochondrial Toxicity Assay in Neuro2A Cells

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Example 5

Mitochondrial Toxicity Assays in Neuro2A Cells

To estimate the potential of the compounds of this invention to cause neuronal toxicity, mouse Neuro2A Cells (American Type Culture Collection 131) can be used as a model system (see Ray A S, Hernandez-Santiago B I, Mathew J S, Murakami E, Bozeman C, Xie M Y, Dutschman G E, Gullen E, Yang Z, Hurwitz S, Cheng Y C, Chu C K, McClure H, Schinazi R F, Anderson K S. Mechanism of anti-human immunodeficiency virus activity of beta-D-6-cyclopropylamino-2′,3′-didehydro-2′,3′-dideoxyguanosine. Antimicrob. Agents Chemother. 2005, 49, 1994-2001). The concentrations necessary to inhibit cell growth by 50% (CC50) can be measured using the 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide dye-based assay, as described. Perturbations in cellular lactic acid and mitochondrial DNA levels at defined concentrations of drug can be carried out as described above. ddC and AZT can be used as control nucleoside analogs.

US10011629B2. Nucleoside analogs for treatment of the flaviviridae family of viruses and cancer (2018-07-03). Cocrystal Pharma, Inc. [US], Emory University [US]. Inventors: Steven J. Coats [US], Franck Amblard [US], Seema Mengshetti [US], Hao Li [US], Raymond F. Schinazi [US].

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Mitochondrial Toxicity Evaluation in Neuro2A Cells

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Example 4

Mitochondrial Toxicity Assays in Neuro2A Cells

To estimate the potential of the compounds of this invention to cause neuronal toxicity, mouse Neuro2A Cells (American Type Culture Collection 131) can be used as a model system (see Ray A S, Hernandez-Santiago B I, Mathew J S, Murakami E, Bozeman C, Xie M Y, Dutschman G E, Gullen E, Yang Z, Hurwitz S, Cheng Y C, Chu C K, McClure H, Schinazi R F, Anderson K S. Mechanism of anti-human immunodeficiency virus activity of beta-D-6-cyclopropylamino-2′,3′-didehydro-2′,3′-dideoxyguanosine. Antimicrob. Agents Chemother. 2005, 49, 1994-2001). The concentrations necessary to inhibit cell growth by 50% (CC₅₀) can be measured using the 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide dye-based assay, as described. Perturbations in cellular lactic acid and mitochondrial DNA levels at defined concentrations of drug can be carried out as described above. ddC and AZT can be used as control nucleoside analogs.

US10752584B2. Elimination of hepatitis B virus with antiviral agents (2020-08-25). EMORY UNIVERSITY [US]. Inventors: Raymond F. Schinazi [US], Sebastien Boucle [US], Franck Amblard [US], Ozkan Sari [US], Leda Bassit [US].

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Evaluating Mitochondrial Toxicity in Neuro2A Cells

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Example 4

Mitochondrial Toxicity Assays in Neuro2A Cells

To estimate the potential of the compounds of this invention to cause neuronal toxicity, mouse Neuro2A Cells (American Type Culture Collection 131) can be used as a model system (see Ray A S, Hernandez-Santiago B I, Mathew J S, Murakami E, Bozeman C, Xie M Y, Dutschman G E, Gullen E, Yang Z, Hurwitz S, Cheng Y C, Chu C K, McClure H, Schinazi R F, Anderson K S. Mechanism of anti-human immunodeficiency virus activity of beta-D-6-cyclopropylamino-2′,3′-didehydro-2′,3′-dideoxyguanosine. Antimicrob. Agents Chemother. 2005, 49, 1994-2001). The concentrations necessary to inhibit cell growth by 50% (CCso) can be measured using the 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide dye-based assay, as described. Perturbations in cellular lactic acid and mitochondrial DNA levels at defined concentrations of drug can be carried out as described above. ddC and AZT can be used as control nucleoside analogs.

US11859014B2. Peptidomimetics for the treatment of Norovirus infection (2024-01-02). EMORY UNIVERSITY [US]. Inventors: Raymond F Schinazi [US], Franck Amblard [US], Ladislau Kovari [US], Peng Liu [US], Shaoman Zhou [US], Benjamin D Kuiper [US], BRadley J Keusch [US].

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Mitochondrial Toxicity Assessment in Neuro2A Cells

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Example 4

Mitochondrial Toxicity Assays in Neuro2A Cells

To estimate the potential of the compounds of this invention to cause neuronal toxicity, mouse Neuro2A Cells (American Type Culture Collection 131) can be used as a model system (see Ray A S, Hernandez-Santiago B I, Mathew J S, Murakami E, Bozeman C, Xie M Y, Dutschman G E, Gullen E, Yang Z, Hurwitz S, Cheng Y C, Chu C K, McClure H, Schinazi R F, Anderson K S. Mechanism of anti-human immunodeficiency virus activity of beta-D-6-cyclopropylamino-2′,3′-didehydro-2′,3′-dideoxyguanosine. Antimicrob. Agents Chemother. 2005, 49, 1994-2001). The concentrations necessary to inhibit cell growth by 50% (CC₅₀) can be measured using the 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide dye-based assay, as described. Perturbations in cellular lactic acid and mitochondrial DNA levels at defined concentrations of drug can be carried out as described above. ddC and AZT can be used as control nucleoside analogs.

US11207370B2. Peptidomimetics for the treatment of coronavirus and picornavirus infections (2021-12-28). Emory University [US]. Inventors: Raymond F. Schinazi [US], Keivan Zandi [US], Franck Amblard [US].

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Cholesterol Depletion Modulates Drug Effects

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Human hepatocellular carcinoma HepG2 cells and mouse neuroblastoma Neuro2a cells were purchased from ATCC (Rockville, MD, USA). Control human fibroblasts were obtained from Coriell Institute for Medical Research (Camden, NJ, USA). HepG2 and human dermal fibroblasts were maintained in DMEM with 10% fetal bovine serum and Neuro2a were maintained in EMEM with 10% fetal bovine serum. To determine the effect of drugs, cells were plated in 96-well plates and incubated at 37 °C in 5% CO₂ for 48 h in presence and absence of different concentrations of ARI, TRZ, and ARI + TRZ. The treatment was performed in cholesterol deficient medium. HepG2 and human fibroblast cultures were grown in DMEM with 10% delipidated fetal bovine serum, and Neuro2a were grown in EMEM plus N2 supplement. At the end point of the incubation, Hoechst dye was added to all wells in the 96-well plate, and the total number of cells was counted using an ImageXpress Pico and cell counting algorithm using CellReporterXpress. After removing the medium, wells were rinsed twice with 1× PBS and then stored at −80 °C for sterol analysis. All samples were analyzed within 2 weeks of freezing.

Balog M., Anderson A.C., Heffer M., Korade Z, & Mirnics K. (2022). Effects of Psychotropic Medication on Somatic Sterol Biosynthesis of Adult Mice. Biomolecules, 12(10), 1535.

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Optimized Lyssavirus Neutralization Assays

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Selected RABVs and non‐RABV lyssaviruses were initially cultured on Neuro‐2A cells (ATCC cat n. CCL‐131) and further adapted on BSR cells (a clone of BHK‐21). Two protocols slightly modified from fluorescent antibody virus neutralization (mFAVN) and from rapid fluorescent foci inhibition (mRFFIT) test (Cliquet et al, 1998; Warrell et al, 2008), respectively, were applied to test the potency of antibodies under study. CVS‐11 working stock was amplified and titrated on either BSR or BHK‐21, according to the neutralization test adopted, RFFIT or FAVN, respectively. In addition, standard FAVN and RFFIT assays were undertaken to assess the potency of tested antibodies against CVS‐11. Briefly, mFAVN assays were based on standard FAVN but were undertaken on BSR cells.

De Benedictis P., Minola A., Rota Nodari E., Aiello R., Zecchin B., Salomoni A., Foglierini M., Agatic G., Vanzetta F., Lavenir R., Lepelletier A., Bentley E., Weiss R., Cattoli G., Capua I., Sallusto F., Wright E., Lanzavecchia A., Bourhy H, & Corti D. (2016). Development of broad‐spectrum human monoclonal antibodies for rabies post‐exposure prophylaxis. EMBO Molecular Medicine, 8(4), 407-421.

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Cell Culture and Transfection Protocol

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Neuro-2a cells (ATCC CCL-131) were maintained in minimum essential medium supplemented with 2 mml-glutamine, 1 mm sodium pyruvate, non-essential amino acids, 10% fetal bovine serum (FBS), 100 units/ml penicillin, and 100 μg/ml streptomycin. COS-7 cells (ATCC CRL-1651) were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% FBS, 100 units/ml penicillin, and 100 μg/ml streptomycin. AML12 cells (ATCC CRL-2254) were cultured in a 1:1 mixture of DMEM and Ham's F-12 medium supplemented with 10% FBS, 100 units/ml penicillin, 100 μg/ml streptomycin, 0.005 mg/ml insulin, 0.005 mg/ml transferrin, 5 ng/ml selenium, and 40 ng/ml dexamethasone. All cells were maintained at 37 °C, 95% humidity, and 5% CO₂. Cells were transfected with Metafectene® (Biontex GmbH, Munich, Germany) according to the manufacturer's instructions and used for experiments 24 h thereafter. Cells were treated with 400 μm oleic acid bound to bovine serum albumin (BSA; essentially fatty acid-free, Sigma-Aldrich), 1 mm 8-CPT-cAMP, or 1 mm 8-CPT-cGMP for 16 h or with Hanks' buffered salt solution for 4 h before being processed for imaging or lipid hydrolase assays.

Heier C., Kien B., Huang F., Eichmann T.O., Xie H., Zechner R, & Chang P.A. (2017). The phospholipase PNPLA7 functions as a lysophosphatidylcholine hydrolase and interacts with lipid droplets through its catalytic domain. The Journal of Biological Chemistry, 292(46), 19087-19098.

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Neuro2A Cell Differentiation Protocol

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Murine neuroblastoma Neuro2A cells (ATCC, Manassas, VA) were maintained in Dulbecco’s modified Eagle’s medium DMEM H-21 (Sigma Aldrich) supplemented with 10% fetal bovine serum (FBS). Medium was removed and replaced by Opti-MEM (Sigma Aldrich) and transfection was performed using Lipofectamine 2000 according to the manufacturer’s protocol (Invitrogen). After 4 hours, medium was replaced by DMEM-10% FBS. Forty-eight hours after transfection, cell differentiation was induced by replacing the medium with DMEM-1% FBS for another 48 hours.

Laguesse S., Morisot N., Shin J.H., Liu F., Adrover M.F., Sakhai S.A., Lopez M.F., Phamluong K., Griffin WC I.I.I., Becker H.C., Bender K.J., Alvarez V.A, & Ron D. (2017). Prosapip1-Dependent Synaptic Adaptations in the Nucleus Accumbens Drive Alcohol Intake, Seeking, and Reward. Neuron, 96(1), 145-159.e8.

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Culturing HEK293T and Neuro-2a Cells

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HEK293T (ATCC, RRID: CVCL_0063) and Neuro-2a cells (ATCC, RRID: CVCL_0470) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) (Thermo Fisher Scientific) containing 10% fetal bovine serum (FBS) (HyClone), 100 IU/ml penicillin (Thermo Fisher Scientific), and 100 μg/ml streptomycin (Thermo Fisher Scientific) in a humidified incubator at 37°C with 5% CO₂. The cell lines used in this study were not further authenticated and not found to be on the list of commonly misidentified cell lines (International Cell Line Authentication Committee).

Tai Y., Gallo N.B., Wang M., Yu J.R, & Van Aelst L. (2019). Axo-axonic Innervation of Neocortical Pyramidal Neurons by GABAergic Chandelier Cells Requires AnkyrinG-associated L1CAM. Neuron, 102(2), 358-372.e9.

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