| BCRJ Code | 0189 |
| Cell Line | Neuro-2a |
| Species | Mus musculus |
| Vulgar Name | Mouse; A Albino |
| Tissue | Brain |
| Cell Type | Neuroblast |
| Morphology | Neuronal And Amoeboid Stem Cells |
| Disease | Neuroblastoma |
| Growth Properties | Adherent |
| Derivation | Derived from a spontaneous tumour in an albino strain A mouse. Cells produce microtubular protein which is believed to play a role in the contractile system giving axoplasmic flow in nerve cells. |
| Applications | This cell line is a suitable transfection host. The cell line has been used for studies on the mechanism of vinblastine precipitation of microtubular protein, the kinetics of GTP binding to isolated protein, the turnover of microtubules in vivo, and the synthesis and assembly of microtubular protein . The World Organization for Animal Health (OIE) uses the cells for routine diagnosis of rabies. |
| Virus Succeptility: | Herpes simplex virus Vesicular stomatitis virus Human poliovirus 1 |
| Products | acetylcholinesterase; tubulin |
| Biosafety | 1 |
| Culture Medium | Dulbecco's Modified Eagle's Medium (DMEM) with 1% non-essential amino acids, 2 mM L-glutamine, 4.5 g/L glucose and 10% of fetal bovine serum, 10%. |
| Subculturing | Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Remove and discard culture medium. Briefly rinse the cell layer with PBS without calcium and magnesium to remove all traces of serum which contains trypsin inhibitor. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C. NOTE: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 12 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 6th edition, published by Alan R. Liss, N.Y., 2010. |
| Subculturing Medium Renewal | 1 to 2 times per week |
| Subculturing Subcultivation Ratio | 1:3 to 1:6 |
| Culture Conditions | Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C |
| Cryopreservation | 95% FBS + 5% DMSO (Dimethyl sulfoxide) |
| Thawing Frozen Cells | SAFETY PRECAUTION:
It is strongly recommended to always wear protective gloves, clothing, and a full-face mask when handling frozen vials. Some vials may leak when submerged in liquid nitrogen, allowing nitrogen to slowly enter the vial. Upon thawing, the conversion of liquid nitrogen back to its gas phase may cause the vial to explode or eject its cap with significant force, creating flying debris.
NOTE: It is important to avoid excessive alkalinity of the medium during cell recovery. To minimize this risk, it is recommended to place the culture vessel containing the growth medium in the incubator for at least 15 minutes before adding the vial contents. This allows the medium to stabilize at its normal pH (7.0 to 7.6). |
| References | Olmsted JB, et al. Isolation of microtubule protein from cultured mouse neuroblastoma cells. Proc. Natl. Acad. Sci. USA 65: 129-136, 1970. PubMed: 5263744 Klebe RJ, Ruddle FH. Neuroblastoma: Cell culture analysis of a differentiating stem cell system. J. Cell Biol. 43: 69A, 1969. Naslavsky N, et al. Characterization of detergent-insoluble complexes containing the cellular prion protein and its scrapie isoform. J. Biol. Chem. 272: 6324-6331, 1997. PubMed: 9045652 Kaneko K, et al. Evidence for protein X binding to a discontinuous epitope on the cellular prion protein during scrapie prion propagation. Proc. Natl. Acad. Sci. USA 94: 10069-10074, 1997. PubMed: 9294164 Maestrini E, et al. A family of transmembrane proteins with homology to the MET-hepatocyte growth factor receptor. Proc. Natl. Acad. Sci. USA 93: 674-678, 1996. PubMed: 8570614 |
| Depositors | Elizabeth Obino Lima, Fundação Instituto Oswaldo Cruz. |
| Cellosaurus | CVCL_0470 |
