B35 (ATCC® CRL-2754)

Organism: Rattus norvegicus, rat  /  Cell Type: neuronal neuroblast; nitrosoethylurea (NEU) induce  /  Tissue: central nervous system (CNS)  /  Disease: neuroblastoma

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Organism Rattus norvegicus, rat
Tissue central nervous system (CNS)
Cell Type neuronal neuroblast; nitrosoethylurea (NEU) induce
Product Format frozen
Morphology neuronal
Culture Properties adherent
Biosafety Level 1
Disease neuroblastoma
Age 4 to 10 months
Strain BD1X
Applications

B35 cells can be stimulated to differentiate in the presence of dibutyryl cyclic AMP (cAMP) or by serum deprivation.

The cells also may be used to study the metabolism and physiology of nervous tissue and the pathology of nervous disorders.

They are easily transfected with plasmid DNA.


Storage Conditions liquid nitrogen vapor phase
Images
Derivation

Rats were inoculated with N-nitrosoethylurea (NEU) 15 days after conception. Tumors found in the central nervous system (CNS) 4 to 10 months after birth were excised, minced, adapted to culture and cloned. RefSchubert D, et al. Clonal cell lines from the rat central nervous system. Nature 249: 224-227, 1974. PubMed: 4151463

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Comments

The cells retain glutamic acid decarboxylase (GAD) and choline acetyltransferase activities; express gamma aminobutyric acid (GABA). The cells are negative for S100 (S-100) protein. RefSchubert D, et al. Clonal cell lines from the rat central nervous system. Nature 249: 224-227, 1974. PubMed: 4151463 The cells are positive for neuron specific enolase. RefVinores SA, et al. Immunoradiometric and immunohistochemical demonstration of neuron-specific enolase in experimental rat gliomas. Cancer Res. 44: 2595-2599, 1984. PubMed: 6722796

A culture submitted to the ATCC in October 2002 was found to be contaminated with mycoplasma. Progeny were cured by a 21-day treatment with BM Cycline. The cells were assayed for mycoplasma, by the Hoechst stain, PCR and the standard culture test, after a six-week period following treatment. All tests were negative.

Complete Growth Medium The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Subculturing Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
Note: Subculture before confluency.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.05% (w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum that 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 pipetting gently.
  5. Add appropriate aliquots of cell suspension to new culture vessels.
  6. Incubate culture vessels at 37°C.
Subcultivation Ratio: 1:4 to 1:6
Medium Renewal: Every 2 to 3 days
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells: a Manual of Basic Technique by R. Ian Freshney, 3th edition, published by Alan R. Liss, N.Y., 1994.
Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
Name of Depositor P Maness
References

Schubert D, et al. Clonal cell lines from the rat central nervous system. Nature 249: 224-227, 1974. PubMed: 4151463

Vinores SA, et al. Immunoradiometric and immunohistochemical demonstration of neuron-specific enolase in experimental rat gliomas. Cancer Res. 44: 2595-2599, 1984. PubMed: 6722796

Otey CA, et al. B35 neuroblastoma cells: an easily transfected, cultured cell model of central nervous system neurons. Methods Cell Biol. : 287-304, 2003. PubMed: 12884695

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

Schubert D, et al. Clonal cell lines from the rat central nervous system. Nature 249: 224-227, 1974. PubMed: 4151463

Vinores SA, et al. Immunoradiometric and immunohistochemical demonstration of neuron-specific enolase in experimental rat gliomas. Cancer Res. 44: 2595-2599, 1984. PubMed: 6722796

Otey CA, et al. B35 neuroblastoma cells: an easily transfected, cultured cell model of central nervous system neurons. Methods Cell Biol. : 287-304, 2003. PubMed: 12884695