Primary culture of lymphoid organ cells and haemocytes of kuruma shrimp, Penaeus japonicus.
A primary cell culture system was developed for the cells of lymphoid organ tissue of kuruma shrimp, Penaeus japonicus. Minced tissues of lymphoid organs were seeded and incubated at 30 degrees C in medium 199 supplemented with 20% foetal bovine serum, a salt mixture and a lactalbumin hydrolysate (0.1 g/l). Fibroblast-like cells and epithelioid-like cells survived for 54 days. Cells did not survive after trypsin, collagenase or hyaluronidase treatment used for cell dissociation. Mitogens (Con A, PHA-P, Pokeweed) and insulin did not enhance cell proliferation. When penaeid rod-shaped DNA virus (PRDV) was inoculated into the lymphoid organ cell culture, a cytopathic effect was observed within 8 days. On the other hand, large granular haemocytes that were fractionated using a Percoll continuous density gradient were not infected with PRDV in vitro within 10 days, which was the longest period of haemocyte maintenance.
Itami
T
T
Department of Applied Aquabiology, National Fisheries University, Shimonoseki, Yamaguchi 759-6595, Japan. itamit@fish-u-ac.jp
Maeda
M
M
Kondo
M
M
Takahashi
Y
Y
D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002470:Cell Survival; D002478:Cells, Cultured; D003470:Culture Media; D004847:Epithelial Cells / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology; D006434:Hemocytes / Q000166:cytology*; D008221:Lymphoid Tissue / Q000166:cytology*; D033561:Penaeidae / Q000166:cytology*; D013997:Time Factors
D003470:Culture Media