R3 [33-10ras3] (ATCC® CRL-2764)

Organism: Rattus norvegicus, rat  /  Cell Type: neuronal Schwann cell; immortalized with SV40 large T antige  / 

Permits and Restrictions

View Permits

Organism Rattus norvegicus, rat
Cell Type neuronal Schwann cell; immortalized with SV40 large T antige
Product Format frozen
Morphology neuronal
Culture Properties adherent
Biosafety Level 2 Cells contain SV40 viral DNA sequences
Applications
At the restrictive temperature, expression of large T antigen was not detected [PubMed: 3049071].
Storage Conditions liquid nitrogen vapor phase
Images
Tumorigenic Yes
Effects
Yes, the cells form colonies in soft agar
Comments
AntigenExp:
SV40-tsA58 large T antigen [PubMed:3049071] Oncogene:
v-Ha-ras[PubMed:3049071] Products:
Myelin-associated glycoprotein (Mag); Protein and mRNA, negative [PubMed: 12210843] Myelin basic protein (Mbp); Protein and mRNA, negative [PubMed: 12210843] Myelin protein zero (Mpz) (Charcot-Marie-Tooth neuropathy 1B); Protein and mRNA, negative [PubMed: 12210843] peripheral myelin protein 22 (Pmp22); protein, negative; mRNA, positive; [PubMed: 12210843] S100 calcium-binding protein, beta (neural) (S100b) protein, positive [PubMed: 12210843]Receptors:
Nerve growth factor receptor (Ngfr) Protein and mRNA, negative
Comments:
Primary cells were infected with the recombinant retrovirus LJ-tsSVLT containing the temperature sensitive tsA58 large SV40 T oncogene and a gene encoding G418 resistance. Clones were selected in the presence of 0.4 mg/ml G418 and super-infected with Zipras 6 containing v-Ha-ras at the permissive temperature of 33C [PubMed:3049071]. The cells proliferate at both the permissive temperature of 33C and the restrictive temperature of 39.5C. However, the growth rate is increased and growth factor requirements are reduced at the permissive temperature [PubMed: 3049071]. At the restrictive temperature, expression of large T antigen was not detected [PubMed: 3049071]. Expression of p21-v-Ha-ras was not detected [PubMed: 3049071].
A culture submitted to the ATCC in December of 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
Protocol:
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (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 33°C to facilitate dispersal.
  4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 33°C.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended
Medium Renewal: Every 2 to 3 days
Cryopreservation
Freeze medium: Complete growth medium, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 33.0°C
Temperature Effects
Restrictive temperature: 39.5°C yes
Permissive temperature: 33°C yes
Name of Depositor PI Patel
References

Hai M, et al. Comparative analysis of Schwann cell lines as model systems for myelin gene transcription studies. J. Neurosci. Res. 69: 497-508, 2002. PubMed: 12210843

Ridley AJ, et al. Ras-mediated cell cycle arrest is altered by nuclear oncogenes to induce Schwann cell transformation. EMBO J. 7: 1635-1645, 1988. PubMed: 3049071

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

Hai M, et al. Comparative analysis of Schwann cell lines as model systems for myelin gene transcription studies. J. Neurosci. Res. 69: 497-508, 2002. PubMed: 12210843

Ridley AJ, et al. Ras-mediated cell cycle arrest is altered by nuclear oncogenes to induce Schwann cell transformation. EMBO J. 7: 1635-1645, 1988. PubMed: 3049071