20B8 (ATCC® CRL-12582™) Organism: Homo sapiens, human / Cell Type: epithelial; somatic hybrid transfected with plasmid pSV2ne General Information Characteristics Culture Method Specifications History Documentation Print Email Share Share this product Facebook Twitter Google+ Permits 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. 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: Remove and discard culture medium. 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. 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. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C. Medium Renewal: Two to three times weekly Cryopreservation Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSOStorage temperature: liquid nitrogen vapor phase Culture Conditions Atmosphere: air, 95%; carbon dioxide (CO2), 5%Temperature: 37.0°C STR Profile Amelogenin: X,YCSF1PO: 10,11,12D13S317: 12,14D16S539: 9D5S818: 8,9,12D7S820: 10,11THO1: 7,9,9.3TPOX: 6,11vWA: 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 Permits 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 Product Sheet Certificate of Analysis MSDS Other Documentation Cell Micrograph References Cho MS, et al. Expression system for factor VIII. US Patent 6,358,703 dated Mar 19 2002