NE-4C细胞
年限： embryo
运输方式： 冻存运输
生长状态： 贴壁生长
品系： C57Bl/Sv129
组织来源： neuroectodermal
数量： 大量
ATCC Number： CRL-2925™
细胞类型： 其他细胞类型
器官来源： 大脑
细胞形态： 其他
是否是肿瘤细胞： 0
物种来源： 小鼠
Designations: NE-4C
NE-4C细胞Depositors: E Madarasz
Biosafety Level: 1
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Mus musculus
Morphology: neuroepithelial
Source: Organ: brain
Tissue: neuroectodermal
Cell type: neural stem cell
Strain: C57Bl/Sv129
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Antigen Expression: Sox-2, Otx-2, En-1
Age: embryo
Comments: NE-4C细胞The neuroepithelial cell lines, NE-4C (CRL-2925) and NE-GFP-4C (CRL-2926) were established from the cerebral vesicles of 9-day-old mouse embryos lacking the functional p53 genes. The cells from both cell lines differentiated to neurons and astrocytes when exposed to retinoic acid. The GFP-transfected clone, NE-GFP-4C, when implanted into the forebrain of *****, new-born, and fetal mice or into the mid- and forebrain vesicles of early chick embryos was capable of developing morphologically differentiated neurons. [PubMed: 9057134, 15246825].
Propagation: ATCC complete growth medium: Minimum essential medium (Eagle) with non-essential amino acids and 4 mM L-glutamine, 90%; fetal bovine serum, 10%
Atmosphere: 5% CO2 in air recommended
Temperature: 37.0°C
Subculturing: Protocol: Volumes used in this protocol are for 75 sq cm flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Note: The culture flasks should be pre-coated with15�g/ml poly-L-lysine (Sigma Cat #P-9155) at least 2 hours in advance.
Remove and discard culture medium.
Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.05% (w/v) Trypsin - 0.53 mM EDTA solution 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: NE-4C细胞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 37C to facilitate dispersal.
Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
Transfer cell suspension to a centrifuge tube and spin at approximately 125 X g for 5 to 10 minutes. Discard supernatant.
Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new poli-L-lysine coated culture vessels. An inoculum of 2 X 10(4) to 4 X 10(4) viable cells/sq. cm is recommended.
Incubate cultures at 37C. Subculture when cell concentration is between 3 X 10(5) and 4 X 10(5) cells/sq. cm.
Subcultivation ratio: A subcultivation ratio of 1:4 to 1:10 is recommended.
Medium renewal: Every 2 to 3 days
Preservation: Freeze medium: Culture medium, 90%; DMSO,10%
Storage temperature: liquid nitrogen vapor phase
Doubling Time: about 12 hours
Related Products: Recommended medium: ATCC 30-2003
Recommended serum: ATCC 30-2020
ATCC CRL-2926
References: 16172636: Schlett K. et al. Retinoic acid induced neural differentiation in a neuroectodermal cell line immortalized by p53 deficiency. J. Neurosci. Res. 15;47(4):405-415 (1997) PubMed: 9057134
16172637: NE-4C细胞Demeter, K et al. Fate of cloned embryonic neuroectodermal cells implanted into the *****, newborn and embryonic forebrain. Exp. Neurol.188(2):254-267 (2004) PubMed: 15246825
