Quality controls to ensure the usability of hiPSCs
(A) Assessment of episome disappearance in hiPSCs. From left to right: Negative ctrl (-), Limit of detection sample (+), hiPSCs that lost the episome (1), hiPSCs that did not lose the episome (2).
(B) Representative picture of a normal hiPSC karyotype showing that no abnormality was acquired during the reprogramming phase.
(C) Full pluripotency was monitored by immunofluoresence staining of pluripotency markers (SOX2, OCT4, Tra-1-60) (in red). Cells were counterstained with DAPI (in blue). Scale bars: 200 μm. Pictures were acquired with inverted microscope Axiovision Observer.Z1.
(D) After differentiation of hiPSC into embryoid bodies (EB), the capacity of hiPSC to form the three embryonic germ layers was assessed by IF staining with one marker per germ layer (green): ectoderm (PAX6), endoderm (AFP) and mesoderm (SMA). Cells were counterstained with DAPI (blue). Scale bars: 200 μm.