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Alternative method for identifying reconstituted cells.

J W Shay, M A Clark

    Proceedings of the National Academy of Sciences of the United States of America
    |January 1, 1980
    PubMed
    Summary
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    Researchers developed a method to create reconstituted cells by fusing nuclear fragments (karyoplasts) with cytoplasmic fragments (cytoplasts). This technique helps study nuclear-cytoplasmic interactions, revealing the nucleus controls tumorigenicity and steroidogenesis.

    Area of Science:

    • Cell Biology
    • Genetics
    • Cancer Research

    Background:

    • Cellular enucleation separates cells into karyoplasts and cytoplasts.
    • Reconstituted cells are valuable for studying nuclear-cytoplasmic interactions.
    • Identifying reconstituted cells requires specific genetic markers.

    Purpose of the Study:

    • To develop and refine techniques for creating and identifying reconstituted cells.
    • To investigate the role of the nucleus in controlling tumorigenicity and steroidogenesis.
    • To utilize specific cell lines (Y-1 and AMT) with distinct genetic markers.

    Main Methods:

    • Cell enucleation using centrifugation and cytochalasin B.
    • Fusion of karyoplasts and cytoplasts using polyethylene glycol.

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  • Development of an identification procedure using genetic markers (BrdUrd(r) and CAP(r)).

    • Main Results:

    • Successfully reconstituted cells from Y-1 karyoplasts and AMT cytoplasts.
    • Extended techniques to isolate cells reconstituted from AMT karyoplasts fused to Y-1 cytoplasts.
    • Demonstrated that the nucleus dictates phenotypic expression of tumorigenicity and steroidogenesis.

    Conclusions:

    • The nucleus (karyoplast) is sufficient to control tumorigenicity and steroidogenesis.
    • The developed identification procedure is crucial for studying reconstituted cells.
    • This research advances understanding of nuclear-cytoplasmic interactions in cell behavior.