| Abstract |
Primary cells cultures from catfish (Clarias batrachus) and snakeheads
(Ophicephalus striatus) were prepared from whole fry and fingerling organ
tissues of the brain, fins, gonad, heart, kidney, liver, skin and spleen.
Four methods were tried: method A wherein explants were placed onto the
surface of 25-cm^2 Primaria flasks (Falcon), allowed to attach for an hour
before addition of Leibovitz medium (L-15) supplemented with 15% fetal
bovine serum (FBS) (L15-L15); Method B wherein explants were inoculated
into 25-cm^2 Primaria flasks (Falcon) already containing L15-15; Method C
which required forcing minced organ sections through a stainless steel
sieve with the aid of a syringe plunger into a Petri dish containing L15-15
medium; and Method D wherein immersed sections of minced tissues to 0.5%
trypsin-EDTA slowly agitated using a magnetic stirrer for one hour at
25 Celsius. Method B was most effective in the establishment of cell
cultures both fish species. Passage numbers of the cells are to date
catfish gonad (CFG) P-56, catfish heart (CFH) P-51, catfish kidney (CFK)
P-7, catfish liver (CFL) P-8, catfish spleen (CFS) P-54, snakehead gonad
(SHG) P-26, snakehead heart (SHH) P-22, snakehead kidney (SHK) P-19,
snakehead liver (SHL) P-49 and snakehead spleen (SHS) P-76. Attempts to
derive primary cell cultures from organ tissues of the brain, fins, skin
and whole fry were unsuccessful. Established cells were fibroblastic. The
cells grew rapidly and became confluent 24 h after seeding at 20 and 25
Celsius. Both SHS and CFS were susceptible to a virus isolated from
EUS-affected fish in the Philippines. The cells were best maintained at 20
Celsius and stored in liquid nitrogen or -70 Celsius.
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