| BCRJ Code | 0417 |
| Cell Line | Grunt Fin (GF) |
| Species | Haemulon sciurus |
| Vulgar Name | Fish |
| Tissue | Fin |
| Morphology | Fibroblast |
| Disease | Normal |
| Growth Properties | Adherent |
| Age/Ethinicity | Adult Year / |
| Derivation | Derived from fin tissue of the adult salt water blue striped Grunt. |
| Virus Succeptility: | Does not support the replication of influenza A, herpes simplex, adeno, polio, dengue and eastern equine encephalitis viruses. |
| Biosafety | 2 |
| Culture Medium | EMEM (HBSS) (Sigma cat no. M5775) + 10mM Hepes (pH 7.3) + 2mM Glutamine +0.23% (w/v) NaHCO3 + 20% FBS (Heat Inactivated)+ 58mM NaCl |
| Subculturing | Split sub-confluent cultures (70-80%) 1:2 to 1:4 using 0.05% trypsin or trypsin/EDTA; 5% CO₂; 20°C. After resuscitation split cells 1:2 to 1:4 and culture for at least one week with 1-2 media changes before shipping. Plating efficiency can be very low after resuscitation and cells may take up to 3 weeks until growth is fully established. Subculture at 80-100% confluency, not below. Cells are sensitive to plastic surface; Falcon / Corning plates and flasks have been used successfully. Fish cell lines detach easily during transit if the culture is too young. |
| Subculturing Medium Renewal | 1 to 2 times per week |
| Subculturing Subcultivation Ratio | 1:2 to 1:4 |
| Culture Conditions | Atmosphere: air, 95%; carbon dioxide (CO2), 5% |
| Cryopreservation | 95% FBS + 5% DMSO (Dimethyl sulfoxide) |
| Thawing Frozen Cells | SAFETY PRECAUTION: Is highly recommend that protective gloves and clothing always be used and a full face mask always be worn when handling frozen vials. It is important to note that some vials leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vessel exploding or blowing off its cap with dangerous force creating flying debris. 1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the Oring and cap out of the water. Thawing should be rapid (approximately 2 minutes). 2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions. 3. For cells that are sensitive to DMSO is recommended that the cryoprotective agent be removed immediately. Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium and spin at approximately 125 x g for 5 to 7 minutes. 4.Discard the supernatant and Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). 5. Incubate the culture in a appropriate atmosphere and temperature (see "Culture Conditions" for this cell line). NOTE: It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6). |
| References | Proc Soc Exp Biol Med 1961;108:762; Ann NY Acad Sci 1965;126;343 |
| Depositors | Lanagro |
| ATCC | CCL-58 |
