Live-Cell Imaging of Human Pluripotent Stem Cells by a Novel Lectin Probe rBC2LCN
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 wordsPluripotent stem cells iPS ES Staining rBC2LCN Lectin
Human iPS cell line 201B7 (HPS0063) was obtained from the RIKEN Bioresource Center.
- 4.Tateno H, Toyoda M, Saito S, Onuma Y, Ito Y, Hiemori K, Fukumura M, Nakasu A, Nakanishi M, Ohnuma K, Akutsu H, Umezawa A, Horimoto K, Hirabayashi J, Asashima M (2011) Glycome diagnosis of human induced pluripotent stem cells using lectin microarray. J Biol Chem 286:20345–20353PubMedCrossRefPubMedCentralGoogle Scholar
- 6.Tateno H, Matsushima A, Hiemori K, Onuma Y, Ito Y, Hasehira K, Nishimura K, Ohtaka M, Takayasu S, Nakanishi M, Ikehara Y, Ohnuma K, Chan T, Toyoda M, Akutsu H, Umezawa A, Asashima M, Hirabayashi J (2013) Podocalyxin is a glycoprotein ligand of the human pluripotent stem cell-specific probe rBC2LCN. Stem Cells Transl Med 2:265–273PubMedCrossRefPubMedCentralGoogle Scholar