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Related Experiment Videos

Lambda-chain production in human lymphoblast-mouse fibroblast hybrids.

S H Orkin, P D Buchanan, W J Yount

    Proceedings of the National Academy of Sciences of the United States of America
    |August 1, 1973
    PubMed
    Summary

    Human lymphoblast cells hybridized with mouse fibroblasts readily produced human lambda chains. This indicates feasibility for gene localization and linkage analysis of immunoglobulin chains using this hybrid cell system.

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    Area of Science:

    • Immunogenetics
    • Cell Biology
    • Molecular Genetics

    Background:

    • Human lymphoblast cells and mouse fibroblasts were used as parental lines.
    • Parental lines were deficient in specific enzymes: hypoxanthine phosphoribosyltransferase (HPRT) and thymidine kinase (TK).
    • Human lymphoblasts produced only the lambda immunoglobulin chain.

    Purpose of the Study:

    • To create and select hybrid cells from human lymphoblasts and mouse fibroblasts.
    • To investigate the stability of differentiated functions, specifically immunoglobulin chain production, in hybrid cells.
    • To assess the feasibility of gene localization and linkage analysis for human immunoglobulin chains.

    Main Methods:

    • Hybridization of HPRT-deficient human lymphoblasts with TK-deficient mouse fibroblasts.

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  • Selection of hybrid cells using hypoxanthanthine-aminopterin-thymidine (HAT) medium.
  • Immunofluorescent techniques to detect immunoglobulin chain production.
  • 5-bromodeoxyuridine selection to assess gene linkage.

    • Main Results:

    • Hybrid cells were successfully selected and maintained production of human lambda chain.
    • No human immunoglobulin heavy chains, complement proteins (C3, C4), or alpha(1)-antitrypsin were detected.
    • Mouse immunoglobulin chains and complement C5 were also absent in hybrids.
    • Selection against TK-positive cells did not affect lambda chain production, suggesting non-linkage of the TK and lambda chain loci.

    Conclusions:

    • Human lymphoblast-mouse fibroblast hybrids can stably produce immunoglobulin chains.
    • This system is a viable model for gene localization and linkage studies of human immunoglobulin chains.
    • The results contrast with the typical extinction of differentiated functions in other hybrid systems.