CHO DP-12 clone#1933 [CHO DP-12, clone#1933 aIL8.92 NB 28605/12] (ATCC® CRL-12444)

Organism: Cricetulus griseus, hamster, Chinese  /  Tissue: ovary  / 

Organism Cricetulus griseus, hamster, Chinese
Tissue ovary
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1
Storage Conditions liquid nitrogen vapor phase
Images
Derivation
The clone#1933 aIL8.92 NB 28605/12 cell line was derived by co-transfecting the CHO cell line DP-12 using Lipofection with the vector p6G4V11N35A.choSD.9 designed to coexpress variable light and heavy regions of the murine 6G4.2.5 monoclonal antibody (ATCC HB-11722). Clones were selected in methotrexate.
Comments

The cells are reported to produce 150 mg/L recombinant human anti-IL-8. Both the clone#1933 (ATCC CRL-12444) and clone#1934 (ATCC CRL-12445) antibodies inhibit IL-8 binding to human neutrophils. They also show equivalent neutralizing capabilities to inhibiting IL-8 mediated human neutrophil chemotaxis.

The mammalian expression plasmid, p6G4V11N35AchoSD.9 (identified as p6G425V11N35A.choSD) is deposited as ATCC 209552.
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: 200 nM methotrexate, 0.002 mg/ml recombinant human insulin, Trace Elements A (Cellgro) to a final concentration of 0.1%, Trace Elements B (Cellgro) to a final concentration of 0.1%, 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.
  1. Remove culture medium. Keep the floating cells to transfer with attached cells..
  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 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.
  6. Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:4 is recommended
Medium Renewal: Two to three times weekly

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, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation
Complete growth medium supplemented with 5% (v/v) DMSO. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.
Culture Conditions
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%
Isotype human IgG1
Name of Depositor Genetech, Inc.
Year of Origin 1933
References

Hsei V, et al. Methods of treating inflammatory diseases with anti-IL-8 antibody fragment-polymer conjugates. US Patent 6,468,532 dated Oct 22 2002

Hsei V, et al. Methods of treating inflammatory disease with anti-IL-8 antibody fragment-polymer conjugates. US Patent 6,458,355 dated Oct 1 2002

Gonzalez TN, et al. Humanized anti-IL-8 monoclonal antibodies. US Patent 6,133,426 dated Oct 17 2000

Gonzalez TN, et al. Humanized anti-IL-8 monoclonal antibodies. US Patent 6,117,980 dated Sep 12 2000

Gonzalez TN, et al. Nucleic acids encoding humanized anti-IL-8 monoclonal antibodies. US Patent 6,025,158 dated Feb 15 2000

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Basic Documentation
Other Documentation
References

Hsei V, et al. Methods of treating inflammatory diseases with anti-IL-8 antibody fragment-polymer conjugates. US Patent 6,468,532 dated Oct 22 2002

Hsei V, et al. Methods of treating inflammatory disease with anti-IL-8 antibody fragment-polymer conjugates. US Patent 6,458,355 dated Oct 1 2002

Gonzalez TN, et al. Humanized anti-IL-8 monoclonal antibodies. US Patent 6,133,426 dated Oct 17 2000

Gonzalez TN, et al. Humanized anti-IL-8 monoclonal antibodies. US Patent 6,117,980 dated Sep 12 2000

Gonzalez TN, et al. Nucleic acids encoding humanized anti-IL-8 monoclonal antibodies. US Patent 6,025,158 dated Feb 15 2000

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.