miPSCs are generated from unpassaged and passaged cultured cells. (A) The miPSC colonies were observed after stable expansion for 30 passages. a, B4 iPSCs; b, C1 iPSCs. (B) The expression of SSEA-1 (green) and Oct4 (green) by miPSCs was detected by immunocytochemistry. (C) Immunohistochemistry for germ layer markers. a, AFP (red, B4 iPSCs); c, AFP (red, C1 iPSCs); e, SMA (green, B4 iPSCs); g, SMA (green, C1 iPSCs); i, Tuj1 (red, B4 iPSCs); k, Tuj1(red, C1 iPSCs); b, d, f, h, j, l, nuclei staining (blue). (D) Teratomas containing multiple tissues are analyzed histologically with Hematoxylin & Eosin staining. a, Neural (derived from B4 iPSCs); b, nerviduct (derived from C1 iPSCs); c, skeletal muscle (derived from B4 iPSCs); d, adipose tissue (derived from C1 iPSCs); e, glandular structures (derived from B4 iPSCs); f, glandular structures (derived from C1 iPSCs). Scale bar, 100 μm.