Neuro2a atcc ccl1 mus musculus brain neuroblastoma. Cytopharmacology of tha is still largely to be discovered. Nov 10, 2004 neuro2a n2a murine neuroblastoma cells are also sensitive to al treatment, which upregulated the expression and accumulation of neurofibrillay tangle protein abreo et al. In this paper we report the effects of the complex iii inhibitor, strobilurin fungicide kresoximmethyl, on the cellular homeostasis of a mammalian cancerous neural cell line. This ability of nb cells to proliferate as well as to.
Mouse neuroblastoma neuro2a n2a cells were plated onto 96. The mouse n2a neuroblastoma cell line was purchased from icn thane, uk. Effect of high ascorbic acid concentrations pharmacological on murine neuroblastoma cell viability. Citeseerx proteomics analysis of the expression of. Every step of the way, a wide range of cell health products. Transfected cells were subsequently cloned, and individual subclones were analysed for their prpc expression levels by immunoblotting using a rabbit polyclonal antibody against aa 94109 of murine prp. A monoclonal antibody called dinutuximab unituxin attaches to gd2, a substance found on the surface of many neuroblastoma cells. Symptoms may include bone pain, a lump in the abdomen, neck, or chest, or a painless bluish lump under the skin. Tacrine 9amino1,2,3,4tetrahydroacridine tha, a potent acetylcholinesterasic inhibitor, is utilized in the pharmacological treatment of alzheimers disease birne and arie, 1994. The differentiated cells have many properties of neurons, including neurofilaments. N2a cells were cultured in mem gibco brl gran island, new york, 14072, usa supplemented with 10% fcs, glutamine, nahco 3 and antibiotics. To give you confidence in the health of your cells every step of the way, weve highlighted the technologies and products within cell biology that are critical to maintaining optimal cell health.
Senitiroh hakomori, kazuko handa, in methods in enzymology, 2003. It was observed that ascorbic acid alone at physiological concentrations 0. Proteomics analysis of the expression of neurogranin in. Strobilurin fungicide kresoximmethyl effects on a cancerous. For this purpose, n2a cells were first stably transfected with. Aug 31, 2017 extracellular fgf1 protects shsy5y cells but not n2a cells from p53dependent apoptosis. A novel epitope for the specific detection of exogenous. Prion propagation in cultured cells british medical. Scrapieinfected murine neuroblastoma cells produce proteaseresistant prion proteins. Neuroblastoma cell n2a culture and cell viability assaysn2a cells were cultured in dulbeccos modified eagles medium supplemented with 10% fetal bovine serum and 2% antibiotic penicillin, erythromycin, and gentamicin in a 5% co 2 atmosphere for 3 days and then transferred to a 96well plate. Neuro2a, a murine neuroblastoma cell line, was purchased from the american type culture collection.
Neuroblastoma nb cell lines are transformed, neural crest derived cells, capable of unlimited proliferation in vitro. Neuroblastoma cell lines center for cancer research. Here we present a proteomics study of murine neuroblastoma n2a cell with lowdose saxitoxin exposure 1 nm and 10 nm, 24h intoxication. Veterinary sciences free fulltext nitric oxide as a. Proteins from neuro2ang cells were labeled with heavy c. In the present paper we report some effects produced by tha on murine neuroblastoma cells n2a used as an experimental model. Here we present a proteomics study of murine neuroblastoma n2a cell. Many neuroblastoma tumor cells will attach to the culture dish, however tumor spheroids may remain in suspension thus the spent media may contain nonadherant tumor cells and should be centrifuged and the pelleted cells recultured in a separate flask. Neuro2a n2a murine neuroblastoma cell lines stably expressing human mutant sod1 g37r 12, 32 were maintained in dmem supplemented with 7% fbs, 2 mm lglutamine, and 400. After selection of n2a clones which carried one of these transactivators, the murine. The activation of exogenous ahr in n2a ralpha cells. A, bar graph of animals dissected at specific time points illustrating the incidence of localized, intermediate, and advanced tumors.
Effects of tacrine upon murine neuroblastoma cells. Induction of neuronal differentiation of murine n2a cells. The mouse neuroblastoma cell line neuro2a ccl1, american type culture collection is grown in dmem. The undifferentiated murine neuroblastoma neuro2a cell line atcc was stably transfected with ahr cdna and the established cell line was named n2a ralpha.
Measles virus induced necrosis in murine neuroblastoma cells. Characterization of detergentinsoluble complexes containing the cellular prion. The human neuroblastoma cell lines shsy5y, imr32, and the mouse cell line neuro2a n2a, derived from a spontaneous neuroblastoma tumour in aj mice, were obtained. Pdf scrapieinfected murine neuroblastoma cells produce. We examined whether exposure to subcytotoxic concentrations of kresoximmethyl induce cellular and biochemical mechanisms of toxicity on the murine neuroblastoma n2a cells. Nbderived gangliosides inhibit the functional activity of t and natural killer cells, contribute to tumorinduced bone marrow suppression, and cause multiple alterations of hematopoiesis, resulting in pancytopenia. Protease omi facilitates neurite outgrowth in mouse neuroblastoma. However, the role of gangliosides in the regulation of. Effects of tacrine upon murine neuroblastoma cells springerlink. The binding of lanthanum to murine neuroblastoma cells clone n1e115 was studied by means of electron microscopy and xray microanalysis.
Using n2a murine neuroblastoma cells, we demonstrated that these cells cultured in the presence of resveratrol and apigenin in a concentration range from 6. In the present study, we used isotopecoded affinity tag icat combined with 2dimensional liquid chromatographytandem mass spectrometry 2dlcmsms to compare the difference of global pattern of protein expression between murine neuroblastoma cells neuro2a and neuro2a cells that stably expressed ng neuro2ang. Pharmaceutical applications of snitrosylation, which, under normal conditions is involved with a host of epigenetic and embryological development pathways, have exhibited great potential for use as adjuvant therapeutics in the clinical management of cancer. B, kaplan meier analysis shows that 65% of mice transgenic for thmycn died of tumor by 95 d. Neuroblastoma cells shsy5y and neuro2a or n2a exposed to atra undergo growth inhibition and neuronal differentiation, both of which are preceded by an increase in clmn mrna.
Neuroblastoma, the most common extracranial solid tumor in children, accounts for 15% of all pediatric cancer deaths. The cells with a starting density of 2x10 4 were treated with ascorbic acid for 24 h. Download scientific diagram mouse neuroblastoma neuro2a n2a cells were plated onto 96well plates, in triplicates, and treated with varying doses of bap. Extracellular fgf1 protects shsy5y cells but not n2a cells from p53dependent apoptosis. These cell lines retain the ability of differentiation into neuronal cell types on treatment with various agents. Neuro2a n2a murine neuroblastoma cells are alsosensitivetoaltreatment,whichupregulatedtheexpression and accumulation of neuro. Programming the lymph node microenvironment to enhance. Given evidence of neuroprotection in animal models of ischemic stroke, we assessed the prenylflavonoid xanthohumol from the common hop humulus lupulus l. Sodium valproate vpa has been reported to increase the accumulation of the pathologic isoform of prion protein prpsc in scrapieinfected murine neuroblastoma cells. M differentiate into mature neurons with dendrites andor axons and that the activation of the pkc signaling pathway plays an important role in this process.
Neuroblastoma is the most common extracranial solid tumor in pediatrics. Combined innate and adaptive immunotherapy overcomes. Several n2a r cell lines were established by transfecting neuro2a cells with the rat ahr cdna, as previously reported. Nbderived gangliosides inhibit the functional activity of t and natural killer cells. Effect of ascorbic acid and hydrogen peroxide on mouse. Neuroblastoma nb, a tumor of the sympathetic nervous system, is the most common extracranial solid tumor in children. Uncovering the proteome response of murine neuroblastoma. In the present paper we report some effects produced by tha on murine neuroblastoma cells n2a. This effect was associated with increased uptake of iron in n2a cells, possibly contributing to alinduced oxidative stress and neurotoxicity. Originating from a mouse, the n2a cell line has a neuronal and amoeboid stem cell morphology, allowing it to differentiate in response to environmental factors. The undifferentiated murine neuroblastoma neuro2a cell line atcc was stably transfected with ahr cdna and the established cell line was named n2ar the activation of exogenous ahr in n2ar. This lanthanum binding was reduced by pharmacological agents that affect muscarinic receptors of these cells or the function of the receptors. Neuroblastoma cell line an overview sciencedirect topics.
Intercellular transmission of a synthetic bacterial. The murine neuroblastoma cell line, n2a, was used to perform a dose response curve for ascorbic acid. Treatment of n2a cells with atra produces a reduction in phosphohistone 3 immunostaining and brdu incorporation, both indicators of a reduction in cell. Cultures may be homogeneous or heterogeneous for these cell types. However, at pharmacological concentrations 16 mm, ascorbic acid caused dosedependent cell.
Prions are infectious, selfpropagating amyloidlike protein aggregates of mammals and fungi. We have studied aggregation propensities of a yeast prion domain in cell culture to gain insights into general mechanisms of prion replication in mammalian cells. It most frequently starts from one of the adrenal glands but can also develop in the neck, chest, abdomen, or spine. It is noteworthy that other neuroblastoma cell lines replicate prion agent with the same efficacy as n2a cells.
Fgf1 protects neuroblastoma shsy5y cells from p53dependent. The toxicity and mode of action of the acute exposure of highdose saxitoxin have been intensively studied for decades. Rna interferencemediated silencing of mutant superoxide. Dyemediated photosensitization of murine neuroblastoma cells. Etoposideinduced apoptosis in murine neuroblastoma n2a. Aluminummaltolate induces apoptosis and necrosis in neuro2a. Cell culture analysis of a differentiating stem cell system. Strikingly, while 75% of mice in the unvaccinated group reached defined endpoints based on tumor burdens by day 15, 75% of mice. To test the putative protective activity of fgf1 on p53dependent cell death in neuroblastoma. Lanthanum binding to murine neuroblastoma cells mayo clinic. Also, the study aimed to investigate the possibility that rf fields might act epipigenetically when combined with xrays, by modifying their effect when applied in vitro to g0 lymphocytes. To assess if the model protein repawh1, and its variants exhibiting distinct aggregation propensities in bacteria 45 47, would show a comparable behavior in mammalian cells, proteins were transiently expressed in murine n2a and human shsy5y neuroblastoma cell lines, commonly used in studies of amyloid neurodegeneration, as well as in.
The mouse neuroblastoma cell line neuro2a ccl1, american type culture collection is grown in dmem supplemented with 10% fcs, 4 m m lglutamine, 1 mm pyruvate, 4. Lanthanum bound only to cellular membranes, in a nonuniform manner. N2a cells are a fastgrowing mouse neuroblastoma cell line. Improved antitumour immunity in murine neuroblastoma using a. The alltrans retinoic acid atraregulated gene calmin. The underlying pathogenic mechanisms of prion infection are not well characterized. Construction and characterization of murine neuroblastoma cell.
Citeseerx studies of genotoxicity and apoptosis using human. Malignant progression and blockade of angiogenesis in a. G0 lymphocytes from 4 donors were exposed for a total of 24 h to a continuous or an. Aluminummaltolate induces apoptosis and necrosis in neuro. Induction of neuronal differentiation of murine n2a cells by. Viologenphosphorus dendrimers exhibit minor toxicity against.
This antibody is now part of the routine treatment for many children with highrisk neuroblastoma, often after a stem cell transplant. In addition, the possibility of rf fields inducing apoptosis in murine neuroblastoma n2a cells. Intercellular transmission of a synthetic bacterial cytotoxic. Treatment of n2a cells with atra produces a reduction in phosphohistone 3 immunostaining and brdu incorporation, both indicators of a reduction in cell proliferation. N2a cells treated with tha at low concentration 1 mu m showed a reduced cells. Maintaining healthy cells is the key to experimental success and reproducible research results. Induction of neuronal differentiation of murine n2a cells by two polyphenols present in the mediterranean diet mimicking neurotrophins activities. Citeseerx document details isaac councill, lee giles, pradeep teregowda. Neuroblastoma nb is a type of cancer that forms in certain types of nerve tissue. Prion protein complexed to n2a cellular rnas through its n. Cytoskeletal protein expression was down regulated in neuro2ang cells. Citeseerx studies of genotoxicity and apoptosis using.
Neuro2a n2a murine neuroblastoma cells are also sensitive to al treatment, which upregulated the expression and accumulation of neurofibrillay tangle protein abreo et al. Neuroblastoma cell n2a culture and cell viability assays n2a cells were cultured in dulbeccos modified eagles medium supplemented with 10% fetal bovine serum and 2% antibiotic penicillin, erythromycin, and gentamicin in a 5% co 2 atmosphere for 3 days and then transferred to a 96well plate. In addition, the possibility of rf fields inducing apoptosis in murine neuroblastoma n2a cells was also examined. G0 lymphocytes from 4 donors were exposed for a total of 24 h to a continuous or an intermittent rf signal. To test the putative protective activity of fgf1 on p53dependent cell death in neuroblastoma models, we. In this study, the effect of vpa on prpsc accumulation was investigated in murine n2a neuroblastoma cells chronically infected with scrapie strain 22l n2a 22l. Mouse neuroblastoma n2a cells and omi proteasedeficient mnd2 mice were used in this study. Mouse n2a cells were grown in dulbeccos modified eagles medium plus 10% fetal bovine serum invitrogen and 2 m m glutamine at 37 c. Murine neuroblastoma n2a cells were purchased from american type culture collection atcc. Compared with normal neuroblastoma cells, n2a cells.
Here, we show that an antiprion antibody derived from camel immunised with murine scrapie material adsorbed to immunomagnetic beads is able to prevent infection of susceptible n2a cells and cure chronically scrapieinfected neuroblastoma cultures. M showed a reduced cell s mitosis and a remarkable reduction of protein synthesis. Mouse neuroblastoma n2a cells were exposed to mouse scrapie prions and. Improved antitumour immunity in murine neuroblastoma using. A camelid antiprp antibody abrogates prpsc replication in. Viologenphosphorus dendrimers exhibit minor toxicity. To study the effect of prion infection on gene expression in neuronal cell cultures, a neuroblastoma n2a cell. This effect was associated with increased uptake of iron in n2a cells. After washing detached cells in media, cells may be resuspended in fresh media in multiple or. N2a cells were cultured in mem gibco brl gran island, new york, 14072, usa supplemented with. Neuroblastomaderived gangliosides inhibit dendritic cell. Alzheimers disease ad is a progressive neurodegenerative disorder that has proved refractory to drug treatment. Differentiation of mouse neuro 2a cells into dopamine neurons. Neuro2a cells produce large quantities of microtubular protein which is believed to play a role in a contractile system which is responsible for axoplasmic flow in.
Neurogranin ng is a neuralspecific, calmodulin cambinding protein that is phosphorylated by protein kinase c pkc. Gene expression profile of a murin neuroblastoma cell line. Construction and characterization of murine neuroblastoma. It is noteworthy that other neuroblastoma cell lines replicate prion agent with the same efficacy as n2a cells table 1 3,9,10,14,18. No accumulation of prpsc was detected after shortterm 3 days. For this purpose, n2a cells were first stably transfected with either the tetracyclinecontrolled transactivator or the reverse transactivator.
Dsmzgermany and the human hela dsmz and neuroblastoma shsy5y cibcsic stock cell lines were cultured in dulbeccos modified eagle. Effect of dioxins on regulation of tyrosine hydroxylase. This antibody can be given together with cytokines immune system hormones such as gmcsf and interleukin2 il2 to help the childs immune system recognize and destroy neuroblastoma cells. The undifferentiated murine neuroblastoma neuro2a cell line atcc was stably transfected with ahr cdna and the established cell line was named n2a r the activation of exogenous ahr in n2a r. Pdf differentiation of n2a cells into dopamine neurons. In addition, to elucidate how conditioned medium exerts neuroprotection, we examined its effect on diazinoninduced neurotoxicity in n2a neuroblastoma cells after it has been incubated with antigdnf antibody. Neuroblastoma cell linesa versatile in vitro model in. Nirenberg by cloning the c0 spontaneous mouse neuroblastoma tumor, c0. N2a cells also known as neuro2a cells are a fastgrowing mouse neuroblastoma cell line. Sodium valproate does not augment prp sc in murine. Expression data from neuro2a mouse neuroblastoma cell lines, anchorage dependent cells ad and anchorageindependent tumorspheres. Two weeks after the boost mice were challenged with 10 6 n2a cells. A tetracyclineinducible expression system has been established for the prion protein prp in murine neuroblastoma cells n2a.
Overall survival is poor for highrisk cases and accounts for about 15% of childhood cancerrelated mortality 1,2,3. The reason for the interest to mouse neuroblastoma n2a cells is that they have a fast growing rate. May, 2003 murine il12 expression by mouse n2a cells was sustained at a fairly constant level throughout the seven days of analysis with no significant difference between single 5306 ng 24 h. Here, we describe the procedure for labeling mouse neuroblastoma cell line neuro2a n2a from atcc. Malignant progression of neuroblastoma in hemizygous mice transgenic for thmycn. The present study aimed to determine whether measles virus can induce apoptosis in murine neuroblastoma cells and the behavior of these cells under acute infection with measles virus or persistent infection with canine distemper virus upon treatment with etoposide.
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