title: Laminin E8 fragments support efficient adhesion and expansion of dissociated human pluripotent stem cells.
abstract: Human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) have the potential to provide an infinite source of tissues for regenerative medicine. Although defined xeno-free media have been developed, culture conditions for reliable propagation of hESCs still require considerable improvement. Here we show that recombinant E8 fragments of laminin isoforms (LM-E8s), which are the minimum fragments conferring integrin-binding activity, promote greater adhesion of hESCs and hiPSCs than do Matrigel and intact laminin isoforms. Furthermore, LM-E8s sustain long-term self-renewal of hESCs and hiPSCs in defined xeno-free media with dissociated cell passaging. We successfully maintained three hESC and two hiPSC lines on LM-E8s in three defined media for 10 passages. hESCs maintained high level expression of pluripotency markers, had a normal karyotype after 30 passages and could differentiate into all three germ layers. This culture system allows robust proliferation of hESCs and hiPSCs for therapeutic applications.
authors.affiliation: Department of Embryonic Stem Cell Research, Institute for Frontier Medical Sciences, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
authors.forename: Takamichi
authors.initials: T
authors.lastname: Miyazaki
authors.forename: Sugiko
authors.initials: S
authors.lastname: Futaki
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Suemori
authors.forename: Yukimasa
authors.initials: Y
authors.lastname: Taniguchi
authors.forename: Masashi
authors.initials: M
authors.lastname: Yamada
authors.forename: Miwa
authors.initials: M
authors.lastname: Kawasaki
authors.forename: Maria
authors.initials: M
authors.lastname: Hayashi
authors.forename: Hideaki
authors.initials: H
authors.lastname: Kumagai
authors.forename: Norio
authors.initials: N
authors.lastname: Nakatsuji
authors.forename: Kiyotoshi
authors.initials: K
authors.lastname: Sekiguchi
authors.forename: Eihachiro
authors.initials: E
authors.lastname: Kawase
mesh_terms: D002448:Cell Adhesion / Q000502:physiology*; D049109:Cell Proliferation; D003470:Culture Media; D005434:Flow Cytometry; D006801:Humans; D007621:Karyotyping; D007797:Laminin / Q000502:physiology*; D039904:Pluripotent Stem Cells / Q000502:physiology*; D020033:Protein Isoforms; D011994:Recombinant Proteins
chemical_list: D003470:Culture Media; D007797:Laminin; D020033:Protein Isoforms; D011994:Recombinant Proteins; C110308:laminin 8