アラマーブルーを用いた稚仔魚の脾臓細胞幼若化の簡便測定法

概要

論文の詳細を見る
A rapid and simple colorimetric assay to determine the blastogenesis of fish larval splenocytes using Alamar Blue was studied. Splenocytes were separated by Percoll gradient from Japanese flounder (one year old), then the blastogenesis of splenocytes stimuiated by mitogen (concanavalin A, pokeweed mitogen, or lipopolysaccharide) as detected using Alamar Blue. The relationship between number of splenocytes and specific absorbance at 72h incubation exhibited a positive, highly significant correlation of p<0.001. Optimum condition of this assay at 5×105cells/well of separated splenocytes with mitogen (concanavalin A 100μg/ml, pokeweed mitogen 10μg/ml, or lipopolysaccaride 1μg/ml) for 72h at 25℃. Splenocytes separation procedure was not required for this blastogenesis assay. Blastogenesis of each splenocyte from larva of Japanese flounder, Japanese parrotfish, and tiger puffer was detectable by this Alamar Blue assay. These results suggested that colorimetric assay using Alamar Blue might be of benefit in a host defense test of cultured fish larva against viral and/or bacterial infections.
長崎大学の論文