Michael Didié, Peter Christalla, Michael Rubart, Vijayakumar Muppala, Stephan Döker, Bernhard Unsöld, Ali El-Armouche, Thomas Rau, Thomas Eschenhagen, Alexander P. Schwoerer, Heimo Ehmke, Udo Schumacher, Sigrid Fuchs, Claudia Lange, Alexander Becker, Wen Tao, John A. Scherschel, Mark H. Soonpaa, Tao Yang, Qiong Lin, Martin Zenke, Dong-Wook Han, Hans R. Schöler, Cornelia Rudolph, Doris Steinemann, Brigitte Schlegelberger, Steve Kattman, Alec Witty, Gordon Keller, Loren J. Field, Wolfram-Hubertus Zimmermann
(A) Undifferentiated PSCs cultured on MEFs formed ESC-like colonies with alkaline phosphatase activity (red – inset). Scale bar: 100 μm. (B) Immunofluorescence labeling of POU5F1, NANOG, and FUT4 (also known as SSEA1) in undifferentiated PSC colonies. Scale bars: 20 μm. (C) Growth kinetics of ESC line R1 and PSC lines A3, A6, B2, and B3 (n = 3 per group and time point; data represent means ± SEM; cell-doubling time: 16–17 hours). (D) PCA of global gene expression profiles of pluripotent cells (PSCs, ESCs, iPSCs, and gPSCs) and somatic cells (MEFs and neural stem cells [NSCs]). The respective Gene Expression Omnibus accession numbers are: GSE11274 (includes MEF and gPSCs); GSE10806 (includes NSC and iPSCs); and GSE30868 (includes ESCs [R1], PSCs [A3] and PSC-derived EBs [A3 EB culture day 15]). Each microarray experiment is represented by a dot, which is positioned in 2D space according to its similarity or degree of variance to all samples analyzed. PC1 and PC2 show variances of 47% and 20%, respectively. (E) Heat map of 80 annotated imprinted genes expressed in ESCs (R1 passage 25; n = 3) and PSCs (A3 passage 25; n = 3). Each gene is represented by a single row of colored boxes. Red represents transcript levels above median; blue represents transcript levels below median. (F) Comparison of global gene expression in ESCs and PSCs with key reprogramming factors highlighted. (G) qPCR of selected stemness factors (n = 4 per group; data represent means ± SEM).