SSN-1

Isolation of fish virus pathogens

Fish striped snakehead fry, spontaneous production of c-type retrovirus

The fish cell line SSN-1 was initiated from whole fry tissue, Channa (Ophicephalus) striatus, commonly named ′striped snakehead′. SSN-1 spontaneously produce and release endogenous snakehead fish Mn²+- dependent C-type retrovirus. The cell line is suitable for fish virus isolation and is susceptible to epizootic ulcerative syndrome rhabdoviruses and piscine nodaviruses. SSN-1 was previously infected with mycoplasma but has been eradicated and tested negative after a minimum of 10 passages.

L15 + 2mM Glutamine + 5-10% Foetal Bovine Serum (FBS).

Split sub-confluent cultures (70-80%) 1:3 to 1:6 i.e. seeding at 2-4x10,000 cells/cm² using 0.25% trypsin or trypsin/EDTA. Use trypsin very cautiously; 25-30°C, can be light sensitive therefore avoid prolonged exposure to bright light. Fish cell lines detach easily during transit if the culture is very young. After resuscitation split cells 1:3 to 1:4 and culture for at least one week with 1-2 media changes before shipping.

When resuscitating a frozen ampoule of cells we recommend the cells are seeded at the higher seeding level i.e. 4 x10, 000/cm². Cells may take up to 10 days to reach 70% following resuscitation and subculture. Therefore media change flasks after 4-5 days to encourage growth.

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