產(chǎn)品名稱 |
UMNSAH/DF-1 |
商品貨號(hào) |
B165910 |
Organism |
Gallus gallus, chicken |
Tissue |
embryo |
Cell Type |
fibroblast spontaneously transformed |
Product Format |
frozen |
Morphology |
fibroblast |
Culture Properties |
adherent |
Biosafety Level |
1
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. |
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 |
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. |
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.
- 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 39°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 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 |
Deposited As |
chicken |
U.S. Patent Number |
|
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
|