RRC ID 45228
Author Tateno H, Onuma Y, Ito Y.
Title Live-cell imaging of human pluripotent stem cells by a novel lectin probe rBC2LCN.
Journal Methods Mol Biol
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.
Volume 1200
Pages 313-8
Published 2014-1-1
DOI 10.1007/978-1-4939-1292-6_26
PMID 25117245
MeSH Carbocyanines / chemistry Cell Differentiation Cell Survival Flow Cytometry Fluorescein-5-isothiocyanate / chemistry Glycomics Humans Lectins / chemistry Lectins / metabolism* Microarray Analysis Molecular Imaging / methods* Molecular Probes / chemistry Molecular Probes / metabolism* Phycoerythrin / chemistry Pluripotent Stem Cells / cytology* Pluripotent Stem Cells / metabolism* Staining and Labeling
Times Cited 4
WOS Category BIOCHEMICAL RESEARCH METHODS BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Human and Animal Cells 201B7(HPS0063)