Methods of preparing an isolated or purified population of thymic emigrant cells and methods of treatment using same

A method of preparing an isolated or purified population of thymic emigrant cells in vitro, comprising:

• modifying T cells that have antigenic specificity for a cancer antigen into pluripotent cells, multipotent cells, or T-lineage cells;
• culturing the pluripotent cells, multipotent cells, or T-lineage cells in the presence of a Notch receptor agonist to produce CD45+ cells;
• culturing the CD45+ cells in the presence of thymic tissue in a hanging drop of medium, wherein culturing the CD45+ cells in the presence of thymic tissue comprises seeding the thymic tissue with the CD45+ cells and migrating the cells into the thymic tissue;
• egressing the cells from the thymic tissue in the hanging drop of medium, wherein the cells egressing from the thymic tissue are thymic emigrant cells and wherein the cells begin to egress from the thymic tissue 2 to 5 days after seeding the thymic tissue with the CD45+ cells;
• and
• isolating the thymic emigrant cells from the thymic tissue without disrupting the thymic tissue, wherein the thymic emigrant cells are CD8α+CD8β+CD4− or CD8α31 CD8β−CD4+, wherein the thymic emigrant cells have the capacity to differentiate into a T cell with a T cell receptor having antigenic specificity for the cancer antigen, and wherein the thymic emigrant cells do not express Ptcra, Rag1, Rag2 and Rorc.