Establishment of BCMA-iPSC from parental BCMA-specific CTL. (A) Schematic presentation of the protocol for reprograming of parental BCMA-specific IFN-γ⁺ CTL into iPSC and differentiation into BCMA-iPSC–T cells. (B, top) Representative flow cytometric analyses showing the pluripotency potential of BCMA-iPSC and EBV-iPSC (positive control) by expression of the stem-cell markers SSEA-4 and TRA-1-60 but not on parental BCMA-CTL (negative control). (B, bottom) Summary of 3 independent analyses (N = 3; mean ± standard deviation [SD]). (C, top) Representative flow cytometric analyses showing the expression of key germ cell layer markers, SOX-17 (endoderm), Brachyury (mesoderm), and Pax-6 (ectoderm), on BCMA-iPSC and EBV-iPSC (positive control). (C, bottom) Summary of 3 independent analyses (N = 3; mean ± SD). (D) Representative immunohistochemistry analysis demonstrating alkaline phosphatase upregulation, a characteristic marker of stem cell development and self-renewal potential, on BCMA-iPSC and EBV-iPSC (positive control) but not on primary T cells. Photomicrographs (original magnification ×100) taken with an inverted microscope (Carl Zeiss). (E) Representative cytogenetic analysis of chromosomes for Giemsa banding patterns demonstrating genomic stability and a normal karyotype in BCMA-iPSC.