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. 2010 Jun 10;20(3):485–493. doi: 10.1089/scd.2010.0128

FIG. 2.

FIG. 2.

Embryonic stem cell (ESC) derivation from Cdx2-deficient embryos and test for pluripotency. (A, B) DIC and fluorescent images of typical outgrowths, 4 days after 8-cell stage embryos had been seeded onto a mouse embryonic fibroblast feeder layer. Cdx2-deficient embryo (B) formed an Oct4-GFP–positive cyst-like structure without the normally forming trophoblast outgrowth in Control (A). Scale bar, 50 μm. (C, D) Ultrastructure of outgrowths on mouse embryonic fibroblasts from 8-cell stage embryos revealed that Cdx2 depletion resulted in an increased number of ESC-like cells (relatively small amount of cytoplasm) inside the outgrowth (D) relative to normal outgrowth (C). Scale bars, 6.5 μm. (E) DIC and fluorescent images showed GFP-positive ESC colonies derived from Cdx2-deficient embryos. Scale bar, 100 μm. (F, H) Chimeras: ESC contribution to somatic tissues of different embryonic germ layers (X-Gal stain-positive in F), germ cells (Oct4-GFP positive in G) in an E13.5 chimera fetus and ESC contribution to columnar intestinal epithelium (X-Gal stain-positive) in an adult chimeric mouse (H). (I–N) Teratoma: cell lines derived from Cdx2-deficient embryos are pluripotent, as evidenced by their ability to form a teratoma (I) with derivatives of all three germ layers, including muscle (mesodermal) (J), gland-like tissue (endodermal) (K), keratinized stratified squamous epithelial cells (ectodermal) (L), osteoid islands showing bony differentiation (mesodermal) (M), and ciliated columnar epithelial cells (endodermal) (N). Color images available online at www.liebertonline.com/scd.