| BCRJ Code | 0307 |
| Cell Line | UMNSAH/DF-1 |
| Species | Gallus gallus |
| Vulgar Name | Chicken |
| Tissue | Embryo |
| Cell Type | Fibroblast Spontaneously Transformed |
| Morphology | Fibroblast |
| Growth Properties | Adherent |
| Age/Ethinicity | 10 (gestation) Day / |
| 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. |
| Applications | The cells are useful as substrates for virus propagation, recombinant protein expression and recombinant virus production |
| Virus Succeptility: | Meleagrid herpesvirus 1 Fowlpox virus Avian reovirus Rous sarcoma virus |
| Tumor Formation: | NO |
| Biosafety | 1 |
| Addtional Info | 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. |
| Culture Medium | Dulbecco's Modified Eagle's Medium (DMEM) modified to contain 2 mM L-glutamine, 4500 mg/L glucose and 10% of fetal bovine serum. |
| 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 PBS without calcium and magnesium 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. NOTE: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 12 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 6th edition, published by Alan R. Liss, N.Y., 2010. |
| Subculturing Medium Renewal | Twice per week |
| Subculturing Subcultivation Ratio | 1:2 to 1:10 is recommended |
| Culture Conditions | Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 39°C; (Max. 40°C, Min. 38°C) |
| Cryopreservation | 95% FBS + 5% DMSO (Dimethyl sulfoxide) |
| Thawing Frozen Cells | SAFETY PRECAUTION:
It is strongly recommended to always wear protective gloves, clothing, and a full-face mask when handling frozen vials. Some vials may leak when submerged in liquid nitrogen, allowing nitrogen to slowly enter the vial. Upon thawing, the conversion of liquid nitrogen back to its gas phase may cause the vial to explode or eject its cap with significant force, creating flying debris.
NOTE: It is important to avoid excessive alkalinity of the medium during cell recovery. To minimize this risk, it is recommended to place the culture vessel containing the growth medium in the incubator for at least 15 minutes before adding the vial contents. This allows the medium to stabilize at its normal pH (7.0 to 7.6). |
| References | Foster DN, Foster LK. Immortalized cell lines for virus growth. US Patent 5,672,485 dated Sep 30 1997 |
| Depositors | MÔNICA MARIA OLIVEIRA PINHO CERQUEIRA - Universidade Federal de Minas Gerais |
| ATCC | CRL-12203 |
| Cellosaurus | CVCL_0570 |
