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Holding medium for cell surface phenotypic analysis.

T J Milson, C W Patrick, N J Torke

    Journal of Immunological Methods
    |March 13, 1986
    PubMed
    Summary
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    This study shows that M199 medium preserves peripheral blood cell surface markers for up to 72 hours. This allows for reliable flow cytometry analysis of B cell lymphoproliferative disease samples after transport.

    Area of Science:

    • Immunology
    • Hematology
    • Biotechnology

    Background:

    • Accurate immunophenotyping of B cell lymphoproliferative disease relies on cell surface marker analysis.
    • Preserving sample integrity during transport and storage is crucial for consistent flow cytometry results.

    Purpose of the Study:

    • To evaluate the efficacy of M199 medium for storing peripheral blood samples for flow cytometry.
    • To determine the stability of cell surface markers on normal and patient samples over 72 hours.

    Main Methods:

    • Peripheral blood samples from healthy adults and B cell lymphoproliferative disease patients were analyzed.
    • Samples were stored in M199 medium (containing insulin, transferrin, L-glutamine) for 0, 24, 48, and 72 hours.
    • Phenotyping was performed using a panel of monoclonal antibodies and flow cytometry (EPICS V).

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    Main Results:

    • No significant differences were observed in cell surface marker expression between initial samples and those stored for up to 72 hours.
    • The whole blood lysis technique proved effective for sample preparation.
    • Flow cytometry analysis demonstrated stable phenotypic profiles across storage durations.

    Conclusions:

    • M199 medium is suitable for the storage and transport of peripheral blood samples for flow cytometry.
    • This method ensures satisfactory cell surface phenotypic analysis for diagnosing B cell lymphoproliferative diseases.
    • The findings support the use of this medium in clinical laboratory settings for sample handling.