Generation of stratified squamous epithelial progenitor cells from mouse induced pluripotent stem cells

• Published in: PloS one 2011-12, Vol.6 (12), p.e28856-e28856
• Main Authors: Yoshida, Satoru, Yasuda, Miyuki, Miyashita, Hideyuki, Ogawa, Yoko, Yoshida, Tetsu, Matsuzaki, Yumi, Tsubota, Kazuo, Okano, Hideyuki, Shimmura, Shigeto
• Format: Article
• Language: English
• Subjects: Animals; Biology; Bone Morphogenetic Protein 4 - pharmacology; Bone morphogenetic proteins; Cell culture; Cell Culture Techniques - methods; Cell Differentiation - drug effects; Cell Proliferation - drug effects; Cells (biology); Cells, Cultured; Clone Cells; Colonies; Cornea - cytology; Developmental biology; Drug screening; E-cadherin; Embryo cells; Embryonic stem cells; Epidermal Cells; Epidermis - drug effects; Epidermis - metabolism; Epithelial cells; Epithelial Cells - cytology; Epithelial Cells - drug effects; Epithelial Cells - ultrastructure; Epithelium; Ethics; Fibroblasts; Genetic disorders; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - drug effects; Induced Pluripotent Stem Cells - metabolism; Medicine; Mice; Physiology; Pluripotency; Polarity; Progenitor cells; Progeny; Regenerative medicine; Rodents; Stem cell transplantation; Stem cells; Stem Cells - cytology; Stem Cells - drug effects; Stem Cells - metabolism; Time Factors; Tissue Engineering; Zonula occludens-1 protein
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0028856

Application of induced pluripotent stem (iPS) cells in regenerative medicine will bypass ethical issues associated with use of embryonic stem cells. In addition, patient-specific IPS cells can be useful to elucidate the pathophysiology of genetic disorders, drug screening, and tailor-made medicine. However, in order to apply iPS cells to mitotic tissue, induction of tissue stem cells that give rise to progeny of the target organ is required. We induced stratified epithelial cells from mouse iPS cells by co-culture with PA6 feeder cells (SDIA-method) with use of BMP4. Clusters of cells positive for the differentiation markers KRT1 or KRT12 were observed in KRT14-positive colonies. We successfully cloned KRT14 and p63 double-positive stratified epithelial progenitor cells from iPS-derived epithelial cells, which formed stratified epithelial sheets consisting of five- to six-polarized epithelial cells in vitro. When these clonal cells were cultured on denuded mouse corneas, a robust stratified epithelial layer was observed with physiological cell polarity including high levels of E-cadherin, p63 and K15 expression in the basal layer and ZO-1 in the superficial layer, recapitulating the apico-basal polarity of the epithelium in vivo. These results suggest that KRT14 and p63 double-positive epithelial progenitor cells can be cloned from iPS cells in order to produce polarized multilayer epithelial cell sheets.