NE-4C (ATCC® CRL-2925)

Organism: Mus musculus, mouse  /  Cell Type: neural stem cell  /  Tissue: brain/neuroectodermal  / 

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Organism Mus musculus, mouse
Tissue brain/neuroectodermal
Cell Type neural stem cell
Product Format frozen
Morphology neuroepithelial
Culture Properties adherent
Biosafety Level 1
Age embryo
Storage Conditions liquid nitrogen vapor phase
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Derivation
The neuroepithelial cell lines, NE-4C (ATCC CRL-2925) and NE-GFP-4C (ATCC CRL-2926) were established from the cerebral vesicles of 9-day-old mouse embryos lacking the functional p53 genes. ­
Antigen Expression
Sox-2, Otx-2, En-1
Comments

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 adult, new-born, and fetal mice or into the mid- and forebrain vesicles of early chick embryos was capable of developing morphologically differentiated neurons. RefSchlett 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 RefDemeter, K et al. Fate of cloned embryonic neuroectodermal cells implanted into the adult, newborn and embryonic forebrain. Exp. Neurol.188(2):254-267 (2004) PubMed: 15246825 


Complete Growth Medium Minimum essential medium (Eagle) with non-essential amino acids and 4 mM L-glutamine, 90%; fetal bovine serum, 10%
Subculturing
Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
Note: The culture flasks should be pre-coated with15µg/mL poly-L-lysine (Sigma Cat #P-9155) at least 2 hours in advance.

  1. Remove and discard culture medium.
  2. 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.
  3. 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Transfer cell suspension to a centrifuge tube and spin at approximately 125 X g for 5 to 10 minutes. Discard supernatant.
  6. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new poly-L-lysine coated culture vessels. An inoculum of 2 X 104 to 4 X 104 viable cells/cm2 is recommended.
  7. Incubate cultures at 37°C. Subculture when cell concentration is between 3 X 105 and 4 X 105 cells/cm2.

Subcultivation ratio: A subcultivation ratio of 1:4 to 1:10 is recommended.
Medium renewal: Every 2 to 3 days.

Cryopreservation
Freeze medium: Culture medium, 90%; DMSO,10%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: 5% CO2 in air recommended
Temperature: 37°C
Population Doubling Time about 12 hours
Name of Depositor E Madarasz
References

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

Demeter, K et al. Fate of cloned embryonic neuroectodermal cells implanted into the adult, newborn and embryonic forebrain. Exp. Neurol.188(2):254-267 (2004) PubMed: 15246825

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation
References

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

Demeter, K et al. Fate of cloned embryonic neuroectodermal cells implanted into the adult, newborn and embryonic forebrain. Exp. Neurol.188(2):254-267 (2004) PubMed: 15246825