title: Efficient Adhesion Culture of Human Pluripotent Stem Cells Using Laminin Fragments in an Uncoated Manner.
abstract: We describe highly effective adhesion culture of human pluripotent stem cells (hPSCs) using laminin fragments without precoating. Culture substrates have been generally thought to exert a cell adhesion effect when they are precoated onto culture vessels. However, simple addition of laminin fragments to a cell suspension during passaging accelerated the adhesion of single dissociated hPSCs onto culture vessels that were not precoated with any culture substrate. Interestingly, similar to conventional precoating, the uncoated addition of laminin fragments supported robust adhesion of single hPSCs and maximum adhesion at a much lower concentration compared with precoating. Similar to precoating laminin fragments, hPSCs seeded with uncoated laminin fragments grew well without cell detachment and maintained pluripotency after continuous subculture. We tested other culture substrates, including full-length laminin and vitronectin, to support hPSC adhesion in the uncoated manner, but only laminin fragments had the potential for application in the uncoated manner. This cost-effective and time-efficient method may contribute to expansion of culture of hPSCs and accelerate the development of regenerative medicine using hPSCs.
authors.affiliation: Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto 6068501, Japan.
authors.forename: Takamichi
authors.initials: T
authors.lastname: Miyazaki
authors.affiliation: Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto 6068501, Japan.
authors.forename: Takehisa
authors.initials: T
authors.lastname: Isobe
authors.affiliation: Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto 6068501, Japan.|Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto 6068507, Japan.
authors.forename: Norio
authors.initials: N
authors.lastname: Nakatsuji
authors.affiliation: Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto 6068507, Japan.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Suemori
mesh_terms: D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D006801:Humans; D007797:Laminin / Q000494:pharmacology*; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects
chemical_list: D007797:Laminin