Rainbow trout leukocyte culture: a simplified method.

Area of Science:

• Aquatic animal health
• Fish immunology
• Cell culture techniques

Background:

• Leukocyte culture is crucial for fish health research.
• Optimizing culture media is essential for successful leukocyte proliferation.
• Previous studies suggested oxygenation is necessary for fish leukocyte growth.

Purpose of the Study:

• To determine optimal conditions for rainbow trout peripheral blood leukocyte culture.
• To evaluate the effect of fetal bovine serum (FBS) concentration on leukocyte growth.
• To investigate the necessity of oxygenation for fish leukocyte culture.

Main Methods:

• Rainbow trout (Salmo gairdneri) peripheral blood leukocytes were cultured in Leibovitz's L-15 medium.
• Cultures were supplemented with varying concentrations of fetal bovine serum (FBS) or homologous serum.
• Cell growth was assessed in both oxygenated and non-oxygenated conditions.

Main Results:

• 10% FBS significantly enhanced peripheral blood leukocyte growth.
• Culture medium without serum or with 20% homologous serum did not support substantial growth.
• Oxygenation of the culture medium was not required for excellent cell growth, contrary to previous reports.

Conclusions:

• Fetal bovine serum at 10% is optimal for culturing rainbow trout leukocytes.
• Successful leukocyte culture can be achieved without medium oxygenation.
• These findings provide a simplified method for fish leukocyte culture, aiding immunological studies.