UMNSAH/DF-1 (ATCC® CRL-12203)

Organism: Gallus gallus, chicken  /  Cell Type: fibroblast spontaneously transformed  /  Tissue: embryo  / 

Organism Gallus gallus, chicken
Tissue
embryo
Cell Type fibroblast spontaneously transformed
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1
Age 10 days gestation
Strain East Lansing Line (ELL-0)
Applications
The cells are useful as substrates for virus propagation, recombinant protein expression and recombinant virus production.
Storage Conditions liquid nitrogen vapor phase
Karyotype Number of cells examined = 59; Modal Chromosome Number = 75 with a range of 65 to 79; Polyploidy Rate = 22%
Derivation
UMNSAH/DF-1 is a spontaneously immortalized chicken cell line derived from 10 day old East Lansing Line (ELL-0) eggs.
Primary chicken embryonic fibroblasts were dissociated and grown in culture; the fibroblasts were passaged until they began to senesce; the cells were concentrated during cell senescence to maintain about 30% to about 60% culture confluence.
Tumorigenic No
Effects
No, in immunosuppressed mice
No, in semisolid medium
Virus Susceptibility Meleagrid herpesvirus 1
Fowlpox virus
Avian reovirus
Rous sarcoma virus
Comments
foci of non-senescent cells were identified and grown for greater than 30 passages.
No clonal proliferation was observed in soft agar cultures, indicating that these cells were immortalized but not transformed.
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: fetal bovine serum to a final concentration of 10%.
Subculturing
Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 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 39°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 39°C.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:10 is recommended
Medium Renewal: Twice per week
Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 39°C; (Max. 40°C, Min. 38°C)
Name of Depositor Regents of the University of Minnesota
Passage History
Primary chicken embryonic fibroblasts were dissociated and grown in culture; the fibroblasts were passaged until they began to senesce; the cells were concentrated during cell senescence to maintain about 30% to about 60% culture confluence;
Foci of non-senescent cells were identified and grown for greater than 30 passages.
References

Foster DN, Foster LK. Immortalized cell lines for virus growth. US Patent 5,672,485 dated Sep 30 1997

Basic Documentation
References

Foster DN, Foster LK. Immortalized cell lines for virus growth. US Patent 5,672,485 dated Sep 30 1997