Materials and methods for the manufacture of pluripotent stem cells

1 . A method of treating a subject in need thereof comprising:

(i) obtaining a population of cells comprising peripheral blood mononuclear cells (PBMCs) from the subject;

(ii) reprogramming γδ T cells in the population of cells to produce iPSCs; and

(iii) administering the produced iPSCs, or a pharmaceutical composition comprising the produced iPSCs to the subject, optionally after differentiating the iPSCs into one or more desired types of cells;

wherein the step of reprogramming γδ T cells comprises:

a) purifying Vγ9Vδ2-enriched γδ T cells from the population of cells;

b) transducing the purified Vγ9Vδ2-enriched γδ T cells with a viral vector encoding one or more reprogramming factors; and

c) 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 Vδ2gene segments;

wherein optionally the subject is a human;

wherein optionally the subject has a hyperproliferative disorder or a cancer of hematopoietic system; and

wherein optionally the viral vector is a Sendai virus (SeV) vector.

2 . The method of claim 1 , wherein the step of purifying the Vγ9Vδ2-enriched γδ T cells from the population of cells comprises further enriching the Vγ9Vδ2-enriched γδ T cells in the isolated population of cells by cell-cell clump enrichment.

3 . The method of claim 1 , wherein the one or more reprogramming factors are selected from the group consisting of OCT3/4, SOX2, KLF4, LIN28, and c-Myc.

4 . The method of claim 1 , wherein in step (b) the transduced cells are cultured in the presence of one or more feeder layers;

wherein optionally in step (b) the transduced cells are cultured in the presence of a mono layer of feeder layer, and

wherein optionally the feeder layer comprises mouse embryonic fibroblasts (MEFs).

5 . The method of claim 1 , wherein the method further comprises isolating the produced iPSCs;

wherein optionally the method further comprises differentiating the iPSCs ex vivo to cells of a desired cell type; and

wherein optionally the method further comprises administering the isolated iPSCs or the differentiated cells to a subject.

6 . The method of claim 1 , wherein the produced iPSCs are negative for a SeV vector.

7 . The method of claim 1 , wherein the produced iPSCs are genomically stable with no loss of a chromosome; wherein optionally the genomic stability of the produced iPSCs is determined by karyotyping analysis.

8 . The method of claim 1 , wherein the produced iPSCs can grow in feeder free medium after adaptation to the feeder free medium.

9 . The method of claim 1 , wherein the viral vector is a SeV vector.