Unit

UNIT 3.14 Production of Mouse T Cell Hybridomas

  1. Ada M. Kruisbeek

Published Online: 1 MAY 2001

DOI: 10.1002/0471142735.im0314s24

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Kruisbeek, A. M. 2001. Production of Mouse T Cell Hybridomas. Current Protocols in Immunology. 24:III:3.14:3.14.1–3.14.11.

Author Information

  1. Netherlands Cancer Institute, Amsterdam, The Netherlands

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: DEC 1997

Abstract

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.