DHFR-G8 (ATCC® CRL-1915)

Organism: Mus musculus, mouse  /  Cell Type: fibroblast fibroblast  / 

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Organism Mus musculus, mouse
Cell Type fibroblast fibroblast
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1
Age embryo
Strain NIH/Swiss
Applications
The DHFR-G8 cell line was produced by M.C. Hung, et al.
This cell line was developed from one methotrexate resistant colony by selection in 0.6 uM methotrexate and 10% dialyzed calf serum.
Storage Conditions liquid nitrogen vapor phase
Derivation
The DHFR-G8 cell line was produced by M.C. Hung, et al., in 1986 from the NIH/3T3 mouse fibroblast cell line which was cotransfected with the cNEU-p clone and the pSV2-DHFR plasmid. This cell line was developed from one methotrexate resistant colony by selection in 0.6 uM methotrexate and 10% dialyzed calf serum. These cells contain 50-100 copies of the cNeu-P (normal rat cosmid DNA) and produce approximately 4 x 10(5) molecules of encoded p185 protein per cell.
Comments
The DHFR-G8 cell line was produced by M.C. Hung, et al., in 1986 from the NIH/3T3 mouse fibroblast cell line which was cotransfected with the cNEU-p clone and the pSV2-DHFR plasmid. This cell line was developed from one methotrexate resistant colony by selection in 0.6 uM methotrexate and 10% dialyzed calf serum. These cells contain 50-100 copies of the cNeu-P (normal rat cosmid DNA) and produce approximately 4 x 10(5) molecules of encoded p185 protein per cell.
Complete Growth Medium Dulbecco's modified Eagle's Medium with 4.5 g/L glucose and 300 nM methotrexate, 90%; bovine calf serum, 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 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 37C 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 37C.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:8 is recommended
Medium Renewal: Every 2 to 3 days
Cryopreservation
Freeze medium: culture medium 95%; DMSO 5%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Name of Depositor RA Weinberg
Year of Origin 1986
References

Hung MC, et al. Molecular cloning of the neu gene: Absence of gross structural alteration in oncogenic alleles. Proc. Natl. Acad. Sci. USA 83: 261-264, 1986. PubMed: 3001730

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
References

Hung MC, et al. Molecular cloning of the neu gene: Absence of gross structural alteration in oncogenic alleles. Proc. Natl. Acad. Sci. USA 83: 261-264, 1986. PubMed: 3001730