title: In vitro modeling of paraxial mesodermal progenitors derived from induced pluripotent stem cells.
abstract: Induced pluripotent stem (iPS) cells are generated from adult somatic cells by transduction of defined factors. Given their unlimited proliferation and differentiation potential, iPS cells represent promising sources for cell therapy and tools for research and drug discovery. However, systems for the directional differentiation of iPS cells toward paraxial mesodermal lineages have not been reported. In the present study, we established a protocol for the differentiation of mouse iPS cells into paraxial mesodermal lineages in serum-free culture. The protocol was dependent on Activin signaling in addition to BMP and Wnt signaling which were previously shown to be effective for mouse ES cell differentiation. Independently of the cell origin, the number of transgenes, or the type of vectors used to generate iPS cells, the use of serum-free monolayer culture stimulated with a combination of BMP4, Activin A, and LiCl enabled preferential promotion of mouse iPS cells to a PDGFR-α(+)/Flk-1(-) population, which represents a paraxial mesodermal lineage. The mouse iPS cell-derived paraxial mesodermal cells exhibited differentiation potential into osteogenic, chondrogenic, and myogenic cells both in vitro and in vivo and contributed to muscle regeneration. Moreover, purification of the PDGFR-α(+)/KDR(-) population after differentiation allowed enrichment of human iPS cell populations with paraxial mesodermal characteristics. The resultant PDGFR-α(+)/KDR(-) population derived from human iPS cells specifically exhibited osteogenic, chondrogenic, and myogenic differentiation potential in vitro, implying generation of paraxial mesodermal progenitors similar to mouse iPS cell-derived progenitors. These findings highlight the potential of protocols based on the serum-free, stepwise induction and purification of paraxial mesodermal cell lineages for use in stem cell therapies to treat diseased bone, cartilage, and muscle.
authors.affiliation: Department of Growth Regulation, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan. hsakurai@cira.kyoto-u.ac.jp
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Sakurai
authors.forename: Yasuko
authors.initials: Y
authors.lastname: Sakaguchi
authors.forename: Emi
authors.initials: E
authors.lastname: Shoji
authors.forename: Tokiko
authors.initials: T
authors.lastname: Nishino
authors.forename: Izumi
authors.initials: I
authors.lastname: Maki
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Sakai
authors.forename: Kazunori
authors.initials: K
authors.lastname: Hanaoka
authors.forename: Akira
authors.initials: A
authors.lastname: Kakizuka
authors.forename: Atsuko
authors.initials: A
authors.lastname: Sehara-Fujisawa
mesh_terms: D028341:Activins / Q000502:physiology; D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D019070:Cell Lineage; D016895:Culture Media, Serum-Free; D015870:Gene Expression / Q000502:physiology; D066298:In Vitro Techniques; D008648:Mesoderm / Q000166:cytology*; D051379:Mice; D008954:Models, Biological; D039904:Pluripotent Stem Cells / Q000166:cytology*; D020796:Receptor, Platelet-Derived Growth Factor alpha / Q000235:genetics / Q000502:physiology; D015398:Signal Transduction; D013234:Stem Cells / Q000166:cytology*; D019076:Transgenes; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000502:physiology; D051153:Wnt Proteins / Q000378:metabolism
chemical_list: C525810:Bmp4 protein, mouse; D055415:Bone Morphogenetic Protein 4; D016895:Culture Media, Serum-Free; D051153:Wnt Proteins; C422405:activin A; D028341:Activins; D020796:Receptor, Platelet-Derived Growth Factor alpha; D040301:Vascular Endothelial Growth Factor Receptor-2