NE-GFP-4C (ATCC® CRL-2926)

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

Permits and Restrictions

View Permits View Restrictions

Organism Mus musculus, mouse
Tissue brain, neuroectodermal
Cell Type neural stem cell
Product Format frozen
Morphology neuroepithelial
Culture Properties adherent
Biosafety Level 1

 


Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Age embryo
Storage Conditions liquid nitrogen vapor phase
Images
Derivation
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.
Antigen Expression
Sox-2, Otx-2, En-1
Comments
Both NE-4C (CRL-2925) and NE-GFP-4C (CRL-2926) differentiate into 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 is 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 The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium:
  • additional 2mM L-Glutamine
    • fetal bovine serum (FBS) to a final concentration of 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 with15 µg/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/cm 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: Complete growth medium,90%; DMSO, 10%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions

Temperature: 37°C

Atmosphere: air, 95%; carbon dioxide (CO2), 5%

Population Doubling Time approximately 16 hours
Name of Depositor E Madarasz
Year of Origin 2000
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
Restrictions

This product's use is governed by the Limited Use License. For information on purchasing a license to use this product for research (for-profit entities) or commercial purposes (any entity) other than those permitted in the limited use label license, contact the Licensing Department, Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, California 92008. Phone (760) 603-7200 or outlicensing@lifetech.com

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