| BCRJ Code | 0368 |
| Cell Line | A-72 |
| Species | Canis familiaris |
| Vulgar Name | Dog |
| Tissue | Unknown |
| Cell Type | Fibroblast |
| Morphology | Fibroblast |
| Disease | Tumor |
| Growth Properties | Adherent |
| Sex | Female |
| Age/Ethinicity | 8 Year / |
| Derivation | The line was established from a 1 cm diameter tumor taken from the left thigh of a female Golden Retriever. |
| Virus Succeptility: | Canine adenovirus 2 Canine coronavirus , Canine coronavirus Canine parainfluenza virus Canid herpesvirus 1 Canine distemper virus Canine minute virus |
| Biosafety | 1 |
| Culture Medium | Leibovitz's L-15 Medium contains 2 mM L-glutamine and no sodium bicarbonate and fetal bovine serum to a final concentration of 10%. Note: The L-15 medium formulation was devised for use in a free gas exchange with atmospheric air. A CO2 and air mixture is detrimental to cells when using this medium for cultivation. |
| Subculturing | Remove medium, rinse with fresh 0.25% trypsin, 0.53 mM EDTA solution, remove trypsin and let the culture sit at 37°C for 10 to 15 minutes. Add fresh medium, aspirate and dispense into new flasks. 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 | 2 to 3 times per week |
| Subculturing Subcultivation Ratio | 1:2 to 1:4 |
| Culture Conditions | Atmosphere: air, 100% Temperature: 37°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 | Binn LN, et al. Establishment of a canine cell line: derivation, characterization, and viral spectrum. Am. J. Vet. Res. 41: 855-860, 1980. PubMed: 6254399 The line was established from a 1 cm diameter tumor taken from the left thigh of a female Golden Retriever. Standard Quantitative Disk Carrier Test Method for Determining the Bactericidal, Virucidal, Fungicidal, Mycobactericidal and Sporicidal Activities of Liquid Chemical Germicides. West Conshohocken, PA:ASTM International;ASTM Standard Test Method E 2197-02. |
| Depositors | Ouro Fino Saúde Animal LTDA |
| Cellosaurus | CVCL_3453 |
