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. Author manuscript; available in PMC: 2014 Apr 15.
Published in final edited form as: Cell Stem Cell. 2008 Sep 11;3(3):340–345. doi: 10.1016/j.stem.2008.08.003

Figure 1. Generation of hiPS cells using inducible lentiviruses.

Figure 1

A. Experimental scheme for the generation of hiPS cells. Primary human fibroblasts and keratinocytes were infected with separate lentiviruses (LV) containing a constitutively active rtTA and doxycycline-inducible reprogramming factors. After infection, cells were seeded to feeders and doxycycline (dox) was applied for 30 days. hiPS clones were picked based on hESC-like morphology and doxycycline-independent growth.

B. Morphology and marker expression in hiPS colonies. Doxycycline-independent fibroblast- and keratinocyte-derived hiPS cells express OCT4 protein.

C. Microarray analysis of gene expression in hiPS cells. Genes with greater than twofold expression level between HUES8 hES cells and BJ fibroblasts were analyzed. Shown are BJ fibroblasts, HUES8 hES cells, and BJ fibroblast-derived hiPS clones.

D. In vitro differentiation of fibroblast-derived hiPS cells into lineages from all three germ layers. Immunostaining for i) Tuj1 (neuronal), ii) cardiac troponin T (cTnT; cardiac muscle), and iii) alpha-fetoprotein (AFP; epithelial, early endodermal).

E. In vitro differentiation of keratinocyte-derived hiPS cells into lineages from all three germ layers. Immunostaining for i) Tuj1, ii) skeletal muscle (MF20), and iii) alpha-fetoprotein.

F. Hematoxylin and eosin stain of teratomas generated from fibroblast-derived hiPS cells. Differentiated structures from all three germ layers were present. i) Pigmented epithelium (ectoderm), ii) cartilage (mesoderm), iii) gut-like epithelium (endoderm), and iv) muscle (mesoderm).