Development of shrimp (Penaeus monodon) cell culture in vitro

Abstract

Primary shrimp cell cultures were successfully developed from lymphoid organ of black tiger shrimp (Penaeus monodon) in double-strength Leibovitz’s L-15 medium supplemented with 15% fetal bovine serum, 1% glucose, 15% shrimp meat extract and 5% lactalbumin hydrolysate. The optimum conditions for primary culture in vitro were obtained in 2 × L-15 medium with an osmolality of 770 mmol/kg at temperature range of 25-28°C. The optimum pH was 7.4. Both epithelial-like and fibroblastic-like cells were observed from lymphoid organ within 2 days incubation. Within three days, 80% confluent monolayers were obtained from lymphoid organ while cultures from other tissues required five days. Cultures were maintained for up to at least 43 days. Only cells from lymphoid organ could be subcultured and confluent monolayers achieved within 10 days post-split. Healthy cultures of the lymphoid cells did not persist beyond the third passage. These primary shrimp cell cultures supported growth of shrimp viruses like WSSV and YHV, and cytopathic effects of these two viruses were characterized.