title: Live-cell imaging of human pluripotent stem cells by a novel lectin probe rBC2LCN.
abstract: We performed comprehensive glycome analysis of a large set of human pluripotent stem cells (hPSCs) using a high-density lectin microarray. We found that a recombinant lectin, rBC2LCN, binds exclusively to all of the undifferentiated hPSCs tested, but not to differentiated somatic cells. rBC2LCN can be used for both the staining and sorting of fixed and live hPSCs. rBC2LCN could serve as a novel detection reagent for hPSCs, particularly given that rBC2LCN is cost effective and, unlike conventional antibodies which require mammalian cells for their production, is easy to produce in a large amount (0.1 g/L) in an Escherichia coli expression system. Here we describe protocols for the fluorescence staining of fixed and live hPSCs and their detection by flow cytometry.
authors.affiliation: Research Center for Stem Cell Engineering, National Institute of Advance Industrial Science and Technology (AIST), Central-2, 1-1-1, Umezono, Tsukuba, Ibaraki, 305-8568, Japan, h-tateno@aist.go.jp.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Tateno
authors.forename: Yasuko
authors.initials: Y
authors.lastname: Onuma
authors.forename: Yuzuru
authors.initials: Y
authors.lastname: Ito
mesh_terms: D002232:Carbocyanines / Q000737:chemistry; D002454:Cell Differentiation; D002470:Cell Survival; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D054794:Glycomics; D006801:Humans; D037102:Lectins / Q000737:chemistry / Q000378:metabolism*; D046228:Microarray Analysis; D057054:Molecular Imaging / Q000379:methods*; D015335:Molecular Probes / Q000737:chemistry / Q000378:metabolism*; D010799:Phycoerythrin / Q000737:chemistry; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D013194:Staining and Labeling
chemical_list: D002232:Carbocyanines; D037102:Lectins; D015335:Molecular Probes; C089187:cyanine dye 3; D010799:Phycoerythrin; D016650:Fluorescein-5-isothiocyanate