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. Author manuscript; available in PMC: 2012 Oct 25.
Published in final edited form as: Curr Biol. 2011 Oct 13;21(20):1759–1765. doi: 10.1016/j.cub.2011.09.010

Figure 4.

Figure 4

ES cell derivation from Blimp1-negative precursors through direct recruitment from epiblast cells. (A) Number of P1 ES cell colonies obtained per embryo. Each symbol represents a single embryo. Bars denote mean values (***P< 0.001, two-tailed t-test). Y-axis is shown in log scale. S, standard conditions; 2i, 2i conditions. (B) Pairwise comparison of global transcriptional profiles between 2i-derived ES cells and R1 ES cells. R, linear coefficient. (C) Histological analysis of teratoma from 2i-derived ES cell lines after injection into SCID mice. (D) Summary of fate-mapping experiments with Blimp1-Cre; Rosa26-RFP embryos. The data are means ± SEM (***P< 0.001, two-tailed t-test). S, standard conditions; 2i, 2i conditions. (E) Representative merged RFP/phase image of an ES cell line (passage 1) derived from Blimp1-Cre; Rosa26-RFP embryo under 2i conditions. Arrowheads indicate RFP+ ES-like colonies. All scale bar = 100 μm. (F) Number of AP+ ES colonies per 100 cells sorted from day-4 ICM outgrowths treated with or without 2i. The data are means ± SEM (*P< 0.05, two-tailed t-test). S, standard conditions; 2i, 2i conditions. (G) Proposed model for generation of ES cells from ICM outgrowth in vitro. Blimp1+ precursors (red) arise from epiblast cells (blue) and differentiate to ES cells in ES cell-derivation medium. These precursors also have the potential to display PGC-like features in the appropriate growth environment. A proportion of epiblast cells not induced toward a germ cell fate can directly acquire ES cell-like characteristics in tissue culture medium supplemented with inhibitors of Erk signaling and Gsk activity (“2i cocktail”; blue line).