Materials and methods for the manufacture of pluripotent stem cells

A method of producing human induced pluripotent stem cells (iPSCs) comprising:

• a) contacting and culturing an isolated population of cells comprising γδ T cells with an activation culture to activate and enrich Vγ9Vδ2 γδ T cells within the isolated population of cells;
• wherein the activation culture comprises IL-15 and zoledronic acid;
• b) after step (a), purifying the Vγ9Vδ2-enriched γδ T cells from the isolated population of cells;
• c) transducing the purified Vγ9Vδ2-enriched γδ T cells with a viral vector encoding one or more reprogramming factors;
• and
• d) culturing the transduced cells under conditions suitable for reprogramming mammalian somatic cells to a pluripotent state, thereby producing iPSCs;
• wherein the produced iPSCs have rearranged TRG and TRD genes;
• wherein the produced iPSCs have TCR rearrangements containing Vγ9 and Vδ2 gene segments;
• wherein optionally the activation culture further comprises IL-2;
• wherein optionally the viral vector is a Sendai virus (SeV) vector;
• wherein optionally the method further comprises obtaining the isolated population of cells from a subject prior to step (a);
• and
• wherein optionally the isolated population of cells are peripheral blood mononuclear cells (PBMCs).