A rapid and simple assay to determine the proliferation of larval and
juveNile fish splenocytes
M.
Yagi, K. Hatasaki, N. Nakahara, K. Hara, K. Tachibana, M. Tsuchimoto-2001
Fish
Pathology, 36(2): 96-98 (from
Current Contents)
Abstract :
A
simple assay to determine the proliferation of larval and juvenile fish
splenocytes using Alamar Blue (AB) was studied. Splenocytes were separated
by Percoll gradient from Japanese flounder Paralichthys olivaceus
(1-year-old), then the proliferation of splenocytes stimulated by mitogen
(concanavalin A: ConA, pokeweed mitogen: PWM, or lipopolysaccharide: LPS)
was detected using AB. The relationship between number of splenocytes and
specific absorbance exhibited a positive significant correlation. The
optimum condition of this assay was 5 x 10(5)cells/well of splenocytes with
mitogens (ConA 100 mug/mL, PWM 10 mug/mL or LPS 1 mug/mL) for 72 h.
Proliferation of each splenocyte from juveniles of Japanese flounder and
larvae of Japanese parrotfish Ophegnathus fasciatus and tiger puffer
Takifugu rubripes was detectable by this assay.
(Nagasaki
Univ, Grad Sch Marine Sci & Engn, Bunkyo Machi 1-14, Nagasaki 8528521,
Japan, e-mail of K. Tachibana: orenge@net.nagasaki-u.ac.jp)