N1E-115 (ATCC® CRL-2263)

Organism: Mus musculus, mouse  /  Cell Type: neuroblast  /  Tissue: brain  /  Disease: neuroblastoma

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Organism Mus musculus, mouse
Tissue brain
Cell Type neuroblast
Product Format frozen
Morphology neuroblast
Culture Properties loosely adherent
Biosafety Level 1
Disease neuroblastoma
Strain A/J
Applications This cell line is a suitable transfection host. The cells can be used to study neurotransmitters and their receptors with radioligand binding, second messenger synthesis and electrophysiological changes.
Storage Conditions liquid nitrogen vapor phase
Karyotype modal number = 192
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Derivation
The N1E-115 cell line was established in 1971 by T. Amano, E. Richelson, and M. Nirenberg by cloning the C-1300 spontaneous mouse neuroblastoma tumor, C-1300.
Receptor Expression
acetylcholine, muscarinic m1; acetylcholine, muscarinic m2; vasoactive intestinal peptide (VIP)
adenosine; angiotensin II; bradykinin; enkephalin; glucagon; histamine H1; 5-hydroxytryptamine (serotonin, 5HT3); neurotensin; prostaglandin E; somatostatin; thrombin
Comments

The clone N1E was subcloned by isolation of single cells on glass shards.

The adrenergic clone N1E-115 exhibits high levels of activity of the enzymes tyrosine hydroxylase and acetylcholinesterase which are necessary for neurotransmitter synthesis but are almost devoid of choline acetyltransferase.

These cells contain 14 receptors for neurotransmitters.


Complete Growth Medium Dulbecco's modified Eagle's medium with 4.5 g/L glucose (without sodium pyruvate), 90%; fetal bovine serum, 10%
Subculturing
  1. Remove medium, rinse with Modified Puck's Saline D1 solution:
    1. Glucose                       5.5 mM
    2. KCl                             5.4 mM
    3. Sucrose                     58.4 mM
    4. Na2HPO4-7H2O         0.17 mM
    5. NaCl                          138 mM
    6. KH2PO4                     0.22 mM
    Adjusted to a pH of 7.4±0.05 with 0.1 N NaOH or 0.N HCl Adjusted to an osmomolality of 340±5 mosM with either sucrose or H20 to decrease or increase the osmomolality.
  2. Allow flasks to remain at room temperature until cells detach.
  3. To remove saline, add fresh culture medium, transfer cell suspension to centrifuge tube and spin at approximately 125 x for 5 to 10 minutes.
  4. Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels.
  5. Place culture vessels in incubators at 37°C

Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:3 is recommended.
Medium Renewal: Every 2 to 3 days.
Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%;if DMEM formulation contain 1.5 g/L sodium bicarbonate.
Temperature: 37°C
Population Doubling Time 36 hrs
Name of Depositor E Richelson
References

. Methods in neurotransmitter receptor analysis. New York: Raven Press; 1990.

Richelson E. The use of cultured cells in the study of mood-normalizing drugs. Pharmacol. Toxicol. 66 suppl.3: 69-75, 1990. PubMed: 2179933

Amano T, et al. Neurotransmitter synthesis by neuroblastoma clones (neuroblast differentiation-cell culture-choline acetyltransferase- acetylcholinesterase-tyrosine hydroxylase-axons-dendrites). Proc. Natl. Acad. Sci. USA 69: 258-263, 1972. PubMed: 4400294

Richelson E. Regulation of tyrosine hydroxylase activity in mouse neuroblastoma clone N1E-115. J. Neurochem. 21: 1139-1145, 1973. PubMed: 4148612

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

. Methods in neurotransmitter receptor analysis. New York: Raven Press; 1990.

Richelson E. The use of cultured cells in the study of mood-normalizing drugs. Pharmacol. Toxicol. 66 suppl.3: 69-75, 1990. PubMed: 2179933

Amano T, et al. Neurotransmitter synthesis by neuroblastoma clones (neuroblast differentiation-cell culture-choline acetyltransferase- acetylcholinesterase-tyrosine hydroxylase-axons-dendrites). Proc. Natl. Acad. Sci. USA 69: 258-263, 1972. PubMed: 4400294

Richelson E. Regulation of tyrosine hydroxylase activity in mouse neuroblastoma clone N1E-115. J. Neurochem. 21: 1139-1145, 1973. PubMed: 4148612