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Fig. 2 | Stem Cell Research & Therapy

Fig. 2

From: Generation of clinical-grade human induced pluripotent stem cells in Xeno-free conditions

Fig. 2

Reprogramming of clinical-grade HFF cells into clinical-grade hiPSCs and characterization of their pluripotency. a Time course of clinical-grade hiPSC generation. b Morphology of clinical-grade HFF cells, and clinical-grade hiPSCs grown on HFF feeder cells and in feeder-free conditions. Scale bar, 200 μm. c Clinical-grade hiPSCs express alkaline phosphatase. Scale bar, 200 μm. d Karyotyping results of one of the clinical-grade hiPSC lines. e mRNA expression levels of pluripotency-related genes of the clinical-grade hiPSCs relative to hESCs. f Immunofluorescence results of one of the clinical-grade hiPSC lines. The hiPSCs express pluripotency markers OCT4, SOX2, SSEA-3, SSEA-4, and TRA-1-60 at the protein level and do not express differentiation marker SSEA-1. Scale bar, 100 μm. g PCR analysis results show that the clinical-grade hiPSCs did not express Sendai virus exogenous genes. h PCR analysis results show that the Sendai virus genes did not integrate into the genomic of the clinical-grade hiPSCs. i Flow cytometry analysis of SSEA-1, SSEA-4, TRA-1-60, and TRA-1-81 expression of one of the clinical-grade hiPSC lines growing without feeders. D day, ESC embryonic stem cell, GMP good manufacturing practice, HFF human foreskin fibroblast, iPSC induced pluripotent stem cell, SSEA stage-specific embryonic antigen

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