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Earthworm coelomocytes in vitro

J Toupin, D H Marks, E L Cooper

    In Vitro
    |April 1, 1977
    PubMed
    Summary
    This summary is machine-generated.

    Researchers developed a method to keep earthworm coelomocytes viable in tissue culture for 10 days. This breakthrough enables in vitro studies of coelomocyte immune responses, overcoming previous contamination and viability issues.

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    Area of Science:

    • Immunology
    • Cell Biology
    • Invertebrate Zoology

    Background:

    • Earthworm coelomocytes are crucial for immune responses, including tissue graft rejection.
    • Previous in vitro studies were hindered by contamination and low cell viability.
    • Developing reliable cell culture methods is essential for studying coelomocyte function.

    Purpose of the Study:

    • To establish a stable in vitro culture method for Lumbricus terrestris coelomocytes.
    • To maintain coelomocyte viability and prevent contamination for extended periods.
    • To enable future in vitro investigations into coelomocyte immune functions.

    Main Methods:

    • Cultured Lumbricus terrestris coelomocytes in L-15 medium with 5-10% fetal bovine serum.
    • Incorporated a cocktail of antibiotics: penicillin, streptomycin, tetracycline, and Amphotericin B.

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  • Maintained cultures at 15°C and assessed viability using trypan blue exclusion and phagocytosis assays.

    • Main Results:

    • Coelomocytes remained viable and uncontaminated for at least 10 days at 15°C.
    • Viability was maintained at 85%, confirmed by trypan blue exclusion and phagocytosis of yeast.
    • Thymidine uptake studies yielded negative results, suggesting limited proliferative capacity under these conditions.

    Conclusions:

    • A robust method for culturing earthworm coelomocytes in vitro has been established.
    • This technique overcomes previous limitations, allowing for extended study of coelomocyte viability.
    • The developed method facilitates in vitro research into the role of coelomocytes in immune responses.