Sialyl-lactotetra, a novel cell surface marker of undifferentiated human pluripotent stem cells.
Cell surface glycoconjugates are used as markers for undifferentiated pluripotent stem cells. Here, antibody binding and mass spectrometry characterization of acid glycosphingolipids isolated from a large number (1 × 10(9) cells) of human embryonic stem cell (hESC) lines allowed identification of several novel acid glycosphingolipids, like the gangliosides sialyl-lactotetraosylceramide and sialyl-globotetraosylceramide, and the sulfated glycosphingolipids sulfatide, sulf-lactosylceramide, and sulf-globopentaosylceramide. A high cell surface expression of sialyl-lactotetra on hESC and human induced pluripotent stem cells (hiPSC) was demonstrated by flow cytometry, immunohistochemistry, and electron microscopy, whereas sulfated glycosphingolipids were only found in intracellular compartments. Immunohistochemistry showed distinct cell surface anti-sialyl-lactotetra staining on all seven hESC lines and three hiPSC lines analyzed, whereas no staining of hESC-derived hepatocyte-like or cardiomyocyte-like cells was obtained. Upon differentiation of hiPSC into hepatocyte-like cells, the sialyl-lactotetra epitope was rapidly down-regulated and not detectable after 14 days. These findings identify sialyl-lactotetra as a promising marker of undifferentiated human pluripotent stem cells.
From the Institute of Clinical Sciences, Department of Surgery, S-41 345 Göteborg, Sweden.
Angela
A
Barone
From the Institute of Clinical Sciences, Department of Surgery, S-41 345 Göteborg, Sweden.
Karin
K
Säljö
the Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology, S-40530 Göteborg, Sweden.
John
J
Benktander
the Institute of Biomedicine, Department of Clinical Chemistry and Transfusion Medicine, S-413 45 Göteborg, Sweden.
Maria
M
Blomqvist
the Institute of Biomedicine, Department of Clinical Chemistry and Transfusion Medicine, S-413 45 Göteborg, Sweden.
Jan-Eric
JE
Månsson
the Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology, S-40530 Göteborg, Sweden.
Bengt R
BR
Johansson
the Institute of Biomedicine, Department of Pathology, S-413 45 Göteborg, Sweden, and.
Johan
J
Mölne
the Cellectis Stem Cells, Cellartis AB, S-413 46 Göteborg, Sweden.
Anders
A
Aspegren
the Cellectis Stem Cells, Cellartis AB, S-413 46 Göteborg, Sweden.
Petter
P
Björquist
From the Institute of Clinical Sciences, Department of Surgery, S-41 345 Göteborg, Sweden, michael.breimer@clinchem.gu.se.
Michael E
ME
Breimer
the Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology, S-40530 Göteborg, Sweden, Susann.Teneberg@medkem.gu.se.
Susann
S
Teneberg
D020384:Acidic Glycosphingolipids / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D015415:Biomarkers / Q000378:metabolism; D002240:Carbohydrate Sequence; D002454:Cell Differentiation; D002460:Cell Line; D015536:Down-Regulation; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D000939:Epitopes / Q000276:immunology; D005434:Flow Cytometry; D005732:Gangliosides / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D013058:Mass Spectrometry; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*
D020384:Acidic Glycosphingolipids; D015415:Biomarkers; D000939:Epitopes; D005732:Gangliosides; C000592361:sialyl-lactotetraosylceramide
Carbohydrate Structure; Cell Surface Immune Recognition; Embryonic Stem Cell; Gangliosides; Glycobiology; Glycolipid Structure; Mass Spectrometry (MS); Sialyl-lactotetraosylceramide; Sulfatide