For expansion and differentiation of human CD34+ hematopoietic progenitor cells to T cells.

Use this two-step protocol to reproducibly expand thousands of CD4+CD8+ double-positive (DP) T cells in cultures initiated with CD34+ human cord blood (CB) cells. In step one, CD34+ CB cells are cultured for 14 days in medium containing expansion supplement to promote their proliferation and differentiation into CD7+CD5+ pro-T cells. In step two, pro-T cells generated during those first 14 days are cultured for another four weeks in medium containing maturation supplement to promote their maturation into DP cells. This medium kit is serum- and feeder-free, eliminating variation introduced by serum and stromal cell lines. For convenience, all kit components are also available for purchase individually.

StemSpan™ lymphoid progenitor expansion supplement (10X) contains a combination of recombinant human cytokines and other additives formulated to selectively promote the expansion and differentiation of CD34+ cells isolated from human CB samples to CD7+CD5+ progenitor T (pro-T) cells. Additionally, StemSpan™ T cell progenitor maturation supplement (10X) enables further maturation of pro-T cells to DP cells. It also supports the maturation of DP cells to CD8 single-positive (SP) T cells with additional stimuli. StemSpan™ lymphoid progenitor expansion supplement (10X) and StemSpan™ T cell progenitor maturation supplement (10X) are intended for use in combination with StemSpan™ SFEM II medium, and on plates coated with StemSpan™ lymphoid differentiation coating material (100X). An optional protocol is available to further mature DP cells to CD8 SP T cells. This extended protocol uses a complete T cell progenitor maturation medium made with reagents included in the StemSpan™ T cell generation kit and must be combined with ImmunoCult™ CD3/CD28/CD2 T cell activator and IL-15.