Production of mouse T cell hybridomas.
T cell hybridomas can be obtained by fusing activated T cells with tumor cells. A heterogeneous population of hybridomas can be cloned by limiting dilution to obtain hybridomas that express specificity to one T cell receptor (TCR). This unit describes cell fusion and selection of T cell hybridomas. A protocol is provided for screening of T cell hybridomas for expression of the CD3-TCR complex by flow cytometry analysis. Those hybridomas expressing a CD3-TCR complex are then further tested for expression of antigen specificity by screening for specific antigen-induced lymphokine production. The procedures for establishment of stable hybridoma lines and cloning of stable lines by limiting dilution are identical to those described for B cell hybridomas and can be found in UNIT 2.5. Protocols for freezing and thawing T cell hybridomas are identical to those described for B cell hybridomas and other cell lines.
Kruisbeek
Ada M
AM
Netherlands Cancer Institute, Amsterdam, The Netherlands.
D000818:Animals; D017252:CD3 Complex / Q000032:analysis / Q000096:biosynthesis; D002459:Cell Fusion; D002999:Clone Cells / Q000276:immunology; D006825:Hybridomas / Q000276:immunology; D007158:Immunologic Techniques; D051379:Mice; D013045:Species Specificity; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism
D017252:CD3 Complex