Preparation of mouse T-cell hybrid tumors
Preparation of mouse T-cell hybrid tumors
T-cell hybridomas can be obtained by fusing activated T cells with tumor cells. Heterogeneous hybridomas can be cloned by limited dilution to obtain hybridomas expressing specific T cell receptors (TCRs). Hybridomas can be expanded in large numbers in the absence of growth factors. The hybridomas proved to be useful for studying T C R-specific recognition of antigens as well as the biochemistry and molecular biology of the C D 3-T C R complex. Author: J.E. Colligan et al, Translated by Xuitao Cao et al. This experiment is from the "Comprehensive Immunology Laboratory Guide".
Operation method
Preparation of mouse T-cell hybrid tumors Move Selection of Basic Solutions Cellular Fusion and T-Cell Hybridomas Materials (see Appendix 1 for items with V) Mouse (for in vivo preparation of soluble protein or peptide sensitized T cells) Complete Freund's adjuvant Reactive cells (unsensitized lymph node T cells) and for activation of T cells in a remixed lymphocyte reaction Stimulated cells (splenocytes expressing the desired alloantigen) B W 5147 lymphoma cell line (A T C C # TIB4 8 , full name B W 5147.G .4.1, drug labeled, dies in H A T medium, resistant to 6-thioguanine) Immediately after the hybridomas have been transferred to a 24-well plate, one well is used for primary screening (e.g. primary screening for C D 3-T C R expression). Selected cell lines should be cloned and frozen by limited dilution (Cell 1.3, Auxiliary Program). 3, Auxiliary Program). In Auxiliary Scheme 1, only a portion of the C D 3-T C R + hybridomas have the required antigenic specificity; therefore, it is necessary to test candidate hybridomas for antigenic specificity rather than to preserve (clone and freeze) all of the C D 3-T C R + hybridomas. For more product details, please visit Aladdin Scientific website.
Ancillary Program 2 Screening for Antigen-Specific Hybridomas 