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. 2016 Jun 2;2:16009. doi: 10.1038/npjparkd.2016.9

Figure 2.

Figure 2

Tests of pluripotency applied to the induced pluripotent stem cells (iPSC) lines generated in the Stem Cell Laboratory of Molecular Brain Research Group, at the University of Eastern Finland (UEF), in Finland. (a) Representative bright field images of human fibroblasts prior to transduction and after the transduction. The putative iPSC colony shows a hES cell-like morphology. (b) The expression levels of Sendai virus in iPSC lines UEF-2B and C (at passage 17; 15), UEF-3B (at passage 15), UEF-4A and B (at passage 17; 15), and UEF-5B, E and F (at passage 18; 15; 16). The levels are compared to non-transduced fibroblast and to hES cells. (c) Representative fluorescence images of iPSC lines stained for the pluripotent markers OCT4, NANOG, TRA1–80 and SSEA4. IPSC colonies stain positive for alkaline phosphatase activity. Images are shown for iPSC line UEF-5F. Scale bars represent 100 μm. (d) Karyogram of iPSC line UEF-2B shows pairs of chromosomes stained using Giemsa (G-banding). (f) IPSC lines form embryoid bodies (EBs) grown in low-adherent plates for 2 weeks (representative image shown for iPSC line UEF-4A, 5B,F). (g) Differentiated EBs generate cells of the three germ layers, immunopositive for alpha-fetoprotein (AFP) (endoderm), smooth muscle antibody (SMA) (mesoderm), and beta III-tubulin (B-III-TUB) (ectoderm); nuclei are counterstained with 4’,6-diamidino-2-2phenylindole (DAPI; shown for iPS lines UEF-3B and UEF-4A). (e) Detection of telomerase activity using TRAPeze telomerase detection kit. (h) Representative images of iPSC lines differentiated towards neural, neuronal, and glial fates. IPSC lines differentiated for 20 days are positive for LMX1A and FOXA2 (midbrain neural progenitors, UEF-1A); when progenitors are kept for two additional weeks in maturation medium, they differentiate into tyrosine hydroxylase (TH)-expressing neurons (shown for iPSC line UEF-4B) that co-label with FOXA2 (shown for iPSC line UEF-2C). Undifferentiated neural progenitors cultured for 4 additional months generate glial fibrillary acidic protein (GFAP)-expressing astrocytes (shown for iPSC line UEF-5B). TH-positive and GFAP-positive cells in a human-derived ventral mesencephalic (hVM) culture served as a reference. Nuclei are counterstained with DAPI. Scale bars represent 100, 50 and 20 μm. (i) Differentiated iPS cells towards tyrosine hydroxylase positive (TH+) cells at day 35. Data are expressed as mean % of TH+ cells compared to DAPI±SD.