A cell culture system developed from heart tissue of the greasy grouper,
284 / 190
Authors
-
K S Sobhana
Marine Biotechnology Division, Central Marine Fisheries Research Institute,
Kochi - 682 018, India
-
K C George
Marine Biotechnology Division, Central Marine Fisheries Research Institute,
Kochi - 682 018, India
-
G Venkat Ravi
Marine Biotechnology Division, Central Marine Fisheries Research Institute,
Kochi - 682 018, India
-
Gijo Ittoop
Marine Biotechnology Division, Central Marine Fisheries Research Institute,
Kochi - 682 018, India
-
R Paulraj
Marine Biotechnology Division, Central Marine Fisheries Research Institute,
Kochi - 682 018, India
Abstract
Heart tissue samples aseptically excised from healthy juveniles of the grouper,
Epinephelus tauvina
were subjected to enzymatic
dissociation using 0.25 % trypsin solution, under aseptic conditions. The resultant cells were suspended in Leibovitz' L-15
medium supplemented with 20 % fetal bovine serum. The cells were subsequently seeded into 25 cm
2
tissue culture flasks at
a density of 10
6 ml-1
and incubated at 28 ± 2°C. Cells showed spreading and attachment to the culture flasks within 24 h and
formed monolayers comprising of epithelioid as well as fibroblast-like cells within 20 days. A confluent monolayer filling
the flask surface comprising predominantly fibroblastic cells was formed within 30 to 35 days. The cell monolayer was
harvested for passage by trypsinisation, which formed successful monolayers in subsequent subcultures.
Â
