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Lectins pp 313-318 | Cite as

Live-Cell Imaging of Human Pluripotent Stem Cells by a Novel Lectin Probe rBC2LCN

  • Hiroaki TatenoEmail author
  • Yasuko Onuma
  • Yuzuru Ito
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1200)

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.

Key words

Pluripotent stem cells iPS ES Staining rBC2LCN Lectin 

Notes

Acknowledgement

Human iPS cell line 201B7 (HPS0063) was obtained from the RIKEN Bioresource Center.

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Research Center for Stem Cell EngineeringNational Institute of Advance Industrial Science and Technology (AIST)TsukubaJapan
  2. 2.Research Center for Stem Cell EngineeringNational Institute of Advance Industrial Science and Technology (AIST)TsukubaJapan

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