FIG. 1.

Reprogramming of PBMCs by SeV vector transduction. (A) Optimization of SeV vector transduction efficiency in peripheral blood cultures using a SeV vector carrying the GFP gene. Representative images of bright field (left panel) and green fluorescence (right panel) were taken at days 1 and 3 after viral transduction. Note the relatively high GFP expression in adherent cells. (B) Schematic protocol of viral transduction. (C) Representative images of iPSC-like colonies from one healthy donor and one CMD patient. The PBMC-derived iPSCs have similar morphology to human embryonic stem cells. (D) Electropherograms of partial sequences of ANKH show mutations maintained in iPSCs: A heterozygous A→G transition in intron 9 leading to an Ala380 insertion, a heterozygous Ser375del, and Phe377del. (E) RT-PCR analysis for the transgenes OCT4, SOX2, c-MYC, KLF4, and SeV vector. HPRT was used as a loading control. Samples were: (1) PBMCs after SeV vector transduction; (2) hiPSCs passage 2; (3) hiPSCs passage 7; (4) hiPSC passage 13; (5) hESC H9. Color images available online at www.liebertpub.com/cell