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20B8 (ATCC® CRL-12582)

Organism: Homo sapiens, human  /  Cell Type: epithelial; somatic hybrid transfected with plasmid pSV2ne  /  Tissue: embryonic kidney  /  Disease: Burkitt's lymphoma

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Organism Homo sapiens, human
Tissue embryonic kidney
Cell Type epithelial; somatic hybrid transfected with plasmid pSV2ne
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Disease Burkitt's lymphoma
Applications
The 20B8 cell line was established by transfection of HKB-11 cells (ATCC CRL-12568) with pCIS25DTR expression vector coding for a B-domain deleted (BDD) FVIII. This cell line can be used for high productivity of B-domain deleted Factor VIII.
Storage Conditions liquid nitrogen vapor phase
Disclosure This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC.
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Derivation
The 20B8 cell line was established by transfection of HKB-11 cells (ATCC CRL-12568) with pCIS25DTR expression vector coding for a B-domain deleted (BDD) FVIII. This cell line can be used for high productivity of B-domain deleted Factor VIII.
Comments
The 20B8 cell line was established by transfection of HKB-11 cells (ATCC CRL-12568) with pCIS25DTR expression vector coding for a B-domain deleted (BDD) FVIII. This cell line can be used for high productivity of B-domain deleted Factor VIII.
Complete Growth Medium The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, ATCC 30-2001. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (ATCC 30-2020) 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 and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM 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.
  6. Incubate cultures 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, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation

Complete growth medium described above 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%
STR Profile
Amelogenin: X,Y
CSF1PO: 10,11,12
D13S317: 12,14
D16S539: 9
D5S818: 8,9,12
D7S820: 10,11
TH01: 7,9,9.3
TPOX: 6,11
vWA: 15,19
Name of Depositor Bayer Corporation
U.S. Patent Number
Disclosure This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC.
References

Cho MS, et al. Expression system for factor VIII. US Patent 6,358,703 dated Mar 19 2002

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.

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

Cho MS, et al. Expression system for factor VIII. US Patent 6,358,703 dated Mar 19 2002

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.