F98npEGFRvIII (ATCC® CRL-2949)

Organism: Rattus norvegicus, rat  /  Cell Type: Glioblastoma  /  Tissue: brain  /  Disease: undifferentiated malignant glioma

Organism Rattus norvegicus, rat
Tissue brain
Cell Type Glioblastoma
Product Format frozen
Morphology glial
Culture Properties adherent
Biosafety Level 2
Disease undifferentiated malignant glioma
Age fetus, 20 days gestation
Applications
This cell line has been used for in vivo studies of molecular targeting of EGFRvIII in syngeneic Fischer rats. RefYang W, et al. Development of a Syngeneic Rat Brain Tumor Model Expressing EGFRvIII and Its Use for Molecular Taregeting Studies with Monoclonal Antibody L8A4. Clin. Cancer Res. 11(1): 341-350, 2005. PubMed: 15671565
Storage Conditions liquid nitrogen vapor phase
Derivation
Epidermal Growth Factor Receptor (EGFR)-expressing F98 glioma cells (F98EGFR) were produced by transfecting F98 (ATCC CRL-2397) with an expression vector containing human EGFRvIII cDNA.
Receptor Expression
Epidermal Growth Factor Receptor vIII (EGFRvIII), expressed (verified at ATCC)
Tumorigenic YES
Effects
tumorigenic in syngeneic Fischer rats
Comments
EGFRvIII is a truncated extracellular mutant of the EGF receptor commonly found in glioblastomas that confers enhanced tumorigenic behavior. The receptors are very weakly phosphorylated. 
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:
  • 0.2 mg/ml G -418
  • fetal bovine serum 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.
    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. Add appropriate aliquots of the cell suspension to new culture vessels.

    Note: If cells are cultured continuously, add 0.6 mg/mL G-418 to the complete growth medium for a passage about once a month.

    Subcultivation ratio: A subcultivation ratio of 1:6 to 1:10 is recommended.

    Medium renewal: Every 2 to 3 days.
    Cryopreservation
    Freeze medium: complete growth medium, 95%; DMSO, 5%
    liquid nitrogen vapor phase
    Culture Conditions
    Temperature: 37°C
    Atmosphere: air, 95%; carbon dioxide (CO2), 5%
    Population Doubling Time about 17 hours
    Name of Depositor RF Barth
    Year of Origin April 2001
    References

    Yang W, et al. Development of a Syngeneic Rat Brain Tumor Model Expressing EGFRvIII and Its Use for Molecular Taregeting Studies with Monoclonal Antibody L8A4. Clin. Cancer Res. 11(1): 341-350, 2005. PubMed: 15671565

    Yang W, et al. Convection enhanced delivery of boronated epidermal growth factor for molecular targeting of EGFR positive gliomas. Cancer Res. 62: 6552-6558, 2002. PubMed: 12438250

    Barth RF, et al. Molecular targeting of the epidermal growth factor receptor for neutron capture therapy of gliomas. Cancer Res. 62(11): 3159-3166, 2002. PubMed: 12036929

    Wu G, et al. Targeted delivery of methotrexate to epidermal growth factor receptor-positive brain tumors by means of cetuximab (IMC-C225) dendrimer bioconjugates. Mol. Cancer Ther. 5(1): 52-59, 2006. PubMed: 16432162.

    Wu G, et al. Molecular targeting and treatment of an epidermal growth factor receptor-positive glioma using boronated cetuximab. Clin. Cancer Res. 13(4): 1260-1268, 2007. PubMed: 17317838

    Yang W, et al. Molecular targeting and treatment of composite EGFR and EGFRvIII-positive gliomas using boronated monoclonal antibodies. Clin. Cancer Res. 14(3): 883-891, 2008. PubMed: 18245552

    Basic Documentation
    References

    Yang W, et al. Development of a Syngeneic Rat Brain Tumor Model Expressing EGFRvIII and Its Use for Molecular Taregeting Studies with Monoclonal Antibody L8A4. Clin. Cancer Res. 11(1): 341-350, 2005. PubMed: 15671565

    Yang W, et al. Convection enhanced delivery of boronated epidermal growth factor for molecular targeting of EGFR positive gliomas. Cancer Res. 62: 6552-6558, 2002. PubMed: 12438250

    Barth RF, et al. Molecular targeting of the epidermal growth factor receptor for neutron capture therapy of gliomas. Cancer Res. 62(11): 3159-3166, 2002. PubMed: 12036929

    Wu G, et al. Targeted delivery of methotrexate to epidermal growth factor receptor-positive brain tumors by means of cetuximab (IMC-C225) dendrimer bioconjugates. Mol. Cancer Ther. 5(1): 52-59, 2006. PubMed: 16432162.

    Wu G, et al. Molecular targeting and treatment of an epidermal growth factor receptor-positive glioma using boronated cetuximab. Clin. Cancer Res. 13(4): 1260-1268, 2007. PubMed: 17317838

    Yang W, et al. Molecular targeting and treatment of composite EGFR and EGFRvIII-positive gliomas using boronated monoclonal antibodies. Clin. Cancer Res. 14(3): 883-891, 2008. PubMed: 18245552