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Related Experiment Videos

A micromethod for evaluating turkey lymphocyte response to phytomitogens.

S K Maheswaran, E S Thies, C Greimann

    American Journal of Veterinary Research
    |September 1, 1975
    PubMed
    Summary

    A novel microculture system enables rapid, reproducible in vitro analysis of turkey lymphocytes. This method optimizes concanavalin A (con A) and pokeweed mitogen stimulation for accurate immune response studies.

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    Area of Science:

    • Immunology
    • Cell Biology
    • Veterinary Science

    Background:

    • Assessing turkey peripheral blood lymphocyte function is crucial for understanding avian immunity.
    • Existing methods for in vitro lymphocyte analysis can be time-consuming and require significant cell numbers.

    Purpose of the Study:

    • To develop and validate a rapid, reproducible microculture system for determining in vitro properties of turkey peripheral blood lymphocytes.
    • To establish optimal conditions for stimulating lymphocytes with concanavalin A (con A) and pokeweed mitogen (PWM).

    Main Methods:

    • A semiautomatic sample harvester was integrated with a microculture system.
    • Turkey peripheral blood lymphocytes were cultured in RPMI 1640 medium without serum.
    • Optimal cell density, mitogen concentration, incubation time, and radiolabeling conditions were determined for con A and PWM stimulation.

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  • Lymphocyte proliferation was measured by 3H-thymidine (3H-TdR) incorporation.

    • Main Results:

    • Optimal con A stimulation required 2 x 10^6 cells/culture, 0.4 µg con A/culture, incubation in flat-bottom wells for 72 hours at 37°C, and 1 µCi 3H-TdR/culture added 12-24 hours before termination.
    • Optimal PWM stimulation conditions were similar, with the exception of using 1 x 10^6 cells/culture in round-bottom wells.
    • PWM yielded maximal stimulation at 80 µg/culture.

    Conclusions:

    • The described microculture system offers a rapid and reproducible method for in vitro lymphocyte analysis in turkeys.
    • Optimized protocols for con A and PWM stimulation provide a reliable basis for further immunological studies in this species.