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. 2016 Mar 30;6:23661. doi: 10.1038/srep23661

Figure 1. Generation and characterization of hiPSCs harboring mtDNA A8344G.

Figure 1

(A) The mtDNA 8344 alanine to guanine mutation rate was quantified by pyrosequencing fibroblasts from patients with MERRF syndrome and MERRF fibroblast-derived hiPSCs after indicated passages (p10–40). (B) Morphology, alkaline phosphatase activity and embryoid body formation of hiPSCs. Scale bar = 50 μm. (C) Reverse transcription polymerase chain reaction (RT-PCR) analysis indicating the expression pattern of embryonic stem cell-like genes in hiPSC sublines. Human embryonic stem cells (hESC) served as a positive control. H2O served as a negative control. (D) Immunofluorescence analysis revealing the protein expression of pluripotency markers (OCT4, NANOG, TRA 1–81 and TRA 1–60) in hiPSC sublines. Nuclei were counterstained with Hoechst 33342. Scale bar = 200 μm. (E) In vitro three-layer differentiation of hiPSCs in specific culture medium showing subpopulations of cells that were immunoreactive for SMA (mesoderm), NESTIN (ectoderm) and AFP (endoderm). Scale bar = 100 μm. (F) Sections of teratomas derived from hiPSC sublines showing the differentiation of histologically distinct tissues, including intestine-, cartilage- and neuron-like tissue. Scale bar = 50 μm. (G) Immunohistochemistry analysis of sections of teratomas derived from hiPSC subline using SMA (mesoderm), AFP (endoderm) and GFAP (ectoderm). Scale bar = 50 μm.