BHK21-pcDNA3.1-HC (ATCC® CRL-13001)

Organism: Mesocricetus auratus, hamster, Syrian golden  /  Cell Type: fibroblast  / 

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Organism Mesocricetus auratus, hamster, Syrian golden
Cell Type fibroblast
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 2 cells contain CMV and SV40 DNA viral particles
Age newborn
Applications
The cells secrete recombinant human erythropoietin in media, which can be detected by ELISA assay.
This line was derived from ATCC PTA-419, which was derived from BHK-21(c-13) (ATCC CCL-10) [U.S. Pat.
Storage Conditions liquid nitrogen vapor phase
Images
Derivation
This line was derived from ATCC PTA-419, which was derived from BHK-21(c-13) (ATCC CCL-10) [U.S. Pat. 6,376,218]. The line was transfected by electroporation using the plasmid vector pcDNA3.1 containing a cDNA fragment encoding human erythropoietin. The vector contains cytomegalovirus (CMV) and SV40 viral DNA sequences and the neomycin resistance gene. The cells secrete recombinant human erythropoietin in media, which can be detected by ELISA assay.
Genes Expressed
recombinant human erythropoietin
Cellular Products
recombinant human erythropoietin
Comments
This line was derived from ATCC PTA-419, which was derived from BHK-21(c-13) (ATCC CCL-10) [U.S. Pat. 6,376,218]. The line was transfected by electroporation using the plasmid vector pcDNA3.1 containing a cDNA fragment encoding human erythropoietin. The vector contains cytomegalovirus (CMV) and SV40 viral DNA sequences and the neomycin resistance gene. The cells secrete recombinant human erythropoietin in media, which can be detected by ELISA assay.
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: 0.4 mg/ml G-418; 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.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended
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
Name of Depositor L Hsu, SC Chang
References

Hsu LW, Chang SC. Expression system for producing recombinant human erythropoietin, method for purifying secreted human erythropoietin and uses thereof. US Patent 6,376,218 dated Apr 28 2002

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

Hsu LW, Chang SC. Expression system for producing recombinant human erythropoietin, method for purifying secreted human erythropoietin and uses thereof. US Patent 6,376,218 dated Apr 28 2002