Description: Immortalized Rat Neural Stem Cells (1464R) are spontaneously immortal population of neural stem cells derived from the brain stem. The cells retained morphology even after 50 passages. In the presence of retinoic acid, the cells may differentiate to TuJ1-positive neurons and to GFAP-positive astrocytes or O4-positive oligodendrocytes. May be used for brain disease studies, and a source for neuronal and glial cells.Immortalization Method: Spontaneous immortalizationBioSafety Level: IISpecies: RatSource Organ: BrainGrowth Properties: AdherentMorphology: PolygonalPopulation Doubling: N/ASeeding Density: N/AMarkers: N/ADonor Age: 12 weeksDonor Gender: N/ADonor Ethnicity: N/APropagation: Cultures are grown on poly(2-hydroxyethylmethacrylate) (PHEMA; Sigma) coated cell culture vessels.
The base medium for this cell line is Neurobasal medium containing 2 mM L-glutamine, 2% B27 supplement (Thermo Fisher Scientific, Waltham, CA, USA), 10 ng/mL of FGF2 (Z101455), 10 ng/mL of EGF (Z100135), and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%.
Change media every 2-3 days.
Carbon dioxide (CO2): 5%, Temperature: +37.0°C.
To differentiate into neuronal and glial cells:
Dissociated cells were seeded, 1-2 x 104 cells per well on poly-L-lysine (PLL)-coated 6 cm dishes. Maintain the cells in differentiation media: F12 medium (Thermo Fisher) supplemented with
fetal bovine serum (TM999)* to a final concentration of 5%, 0.5% N2 supplement (Thermo Fisher), 1% B27 supplement,1 μM all-trans retinoic acid (ATRA; Sigma), and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%.
Change media every 2-3 days.
Carbon dioxide (CO2): 5%, Temperature: +37.0°C.
* Do not use heat-inactivated FBS for cell culture unless specified otherwise.Quality Control: N/AShipping Condition: Dry IceStorage Condition: liquid nitrogen or -180°C