title: Human and mouse induced pluripotent stem cells are differentially reprogrammed in response to kinase inhibitors.
abstract: Conventional human induced pluripotent stem cells (hiPSCs), reprogrammed from somatic cells by induced expression of Oct4, Sox2, Klf4, and c-Myc, are phenotypically different from mouse embryonic stem cells (ESCs). In mice, culture in N2B27 serum-free 2i media (mitogen-activated protein kinase/extracellular signal-regulated kinase and glycogen synthase kinase 3 inhibitors; PD0325901 and CHIR99021) plus leukemia inhibitory factor (LIF) (2i+LIF medium) enriches for germline competent ESCs. Here, we demonstrate that flat-shaped hiPSC colonies can be reprogrammed into bowl-shaped multi-potent stem cells (2i-hiPSCs) by using 2i+LIF medium. Mechanical dissociation of 2i-hiPSC colonies enables stable maintenance for >20 passages. Importantly, gene expression profiling demonstrated that 2i-hiPSCs more closely resemble primitive neural stem cells (PNSCs). Notably, this 2i-induced phenotype was generated from conventional hiPSCs, but not human ESCs (hESCs), thus correlating with the observation of neuroectodermal SOX1-positive colonies in conventional hiPSCs, but not hESCs in 2i+LIF medium. Thus, 2i-hiPSCs, which are nonteratoma forming PNSCs, may represent a safe source of cells for neural research and regenerative medicine.
authors: Laboratory of Stem Cell Engineering, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan.
authors: Kunio
authors: K
authors: Hirano
authors: Shogo
authors: S
authors: Nagata
authors: Shinpei
authors: S
authors: Yamaguchi
authors: Masato
authors: M
authors: Nakagawa
authors: Keisuke
authors: K
authors: Okita
authors: Hidetoshi
authors: H
authors: Kotera
authors: Justin
authors: J
authors: Ainscough
authors: Takashi
authors: T
authors: Tada
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D019070:Cell Lineage / Q000187:drug effects; D065150:Cellular Reprogramming / Q000187:drug effects*; D003470:Culture Media / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology*; D000090062:Kruppel-Like Factor 4; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology; D051379:Mice; D008954:Models, Biological; D039902:Multipotent Stem Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*
chemical_list: D003470:Culture Media; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D053605:Leukemia Inhibitory Factor; D047428:Protein Kinase Inhibitors