20B8 (ATCC® CRL-12582)

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

Organism Homo sapiens, human
Cell Type epithelial; somatic hybrid transfected with plasmid pSV2ne
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2
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
Images
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, Catalog No. 30-2001. 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 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.
Medium Renewal: Two to three times weekly
Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
STR Profile
Amelogenin: X,Y
CSF1PO: 10,11,12
D13S317: 12,14
D16S539: 9
D5S818: 8,9,12
D7S820: 10,11
THO1: 7,9,9.3
TPOX: 6,11
vWA: 15,19
Name of Depositor Bayer Corporation
References

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

Basic Documentation
Other Documentation
References

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