Methods for the isolation and expansion of cord blood derived T regulatory cells

摘要

The present invention encompasses methods, and kits for the isolation and expansion of T regulatory cells having the CD45RA+ phenotype, including such cells from human umbilical cord blood.

主权项

1. A method of generating a regulatory T cell (Treg cell) suppressor cell line, wherein said Treg suppressor cell line suppresses T cell proliferation in a mixed lymphocyte reaction by a factor of at least 95% at a ratio of 1:32 (Treg: T cell), said method comprising:
a) isolating a population of mononuclear cells from a human umbilical cord blood sample; b) contacting said population of mononuclear cells with an antibody that specifically binds CD25 under conditions suitable for formation of a mononuclear cell-antibody complex; c) substantially separating said mononuclear cell-antibody complex from said population of mononuclear cells; thereby isolating a population of phenotypically CD25+CD45RA+ blood cells; d) expanding said isolated population of CD25+ cells in the presence of anti-CD3/CD28 antibody coated beads; e) activating said expanded population of CD25+ cells in the presence of IL-2; and f) substantially separating said activated population of CD25+ cells; thereby generating a suppressor cell line that suppresses T cell proliferation in a mixed lymphocyte reaction by at least 95% when at a ratio of 1:32 (Treg: T cell).