Fig. 5.

Spontaneous and guided differentiation of the MV4F^N-derived induced pluripotent stem cell (iPSC) clones. a Representative MV4F^N-derived iPSC clones obtained with or without small molecules (+sm, −sm) and one 4LV-derived iPSC control (4LV) were analyzed by immunofluorescence for lineage markers for three germ layers (endoderm, mesoderm, and ectoderm). iPSC clones were spontaneously differentiated through embryoid body formation. Pluripotency of derived iPSC clones was verified by generation of cells of ectoderm (β-III tubulin, green, top row), endoderm (FOXA2, red, second row), and mesoderm (CD31, green, bottom row) upon spontaneous differentiation. Clones were tested at passages 4 and 20. Scale bars indicate 50 μm. b Representative MV4F^N-derived iPSC clones obtained with or without small molecules (+sm, −sm), and one 4LV-derived iPSC control (4LV) were analyzed by immunofluorescence for lineage markers for three germ layers (endoderm, mesoderm, and ectoderm). iPSC clones were differentiated through guided differentiation using the STEMdiff™ Trilineage Differentiation kit. Pluripotency of derived iPSC clones was verified by generation of cells of ectoderm (Nestin [red] and PAX-6 [green], top row), endoderm (FOXA2 [red] and SOX17 [green], second row), and mesoderm (NCAM [green] and Brachyury [red], bottom row) upon guided differentiation. Clones were tested at passage 4. Control staining was done on not differentiated iPSCs (right three panels, iPSCs). Scale bars indicate 50 μm