title: Structural complexity of non-acid glycosphingolipids in human embryonic stem cells grown under feeder-free conditions.
abstract: Due to their pluripotency and growth capability, there are great expectations for human embryonic stem cells, both as a resource for functional studies of early human development and as a renewable source of cells for use in regenerative medicine and transplantation. However, to bring human embryonic stem cells into clinical applications, their cell surface antigen expression and its chemical structural complexity have to be defined. In the present study, total non-acid glycosphingolipid fractions were isolated from two human embryonic stem cell lines (SA121 and SA181) originating from leftover in vitro fertilized human embryos, using large amounts of starting material (1 × 10(9) cells/cell line). The total non-acid glycosphingolipid fractions were characterized by antibody and lectin binding, mass spectrometry, and proton NMR. In addition to the globo-series and type 1 core chain glycosphingolipids previously described in human embryonic stem cells, a number of type 2 core chain glycosphingolipids (neo-lactotetraosylceramide, the H type 2 pentaosylceramide, the Le(x) pentaosylceramide, and the Le(y) hexaosylceramide) were identified as well as the blood group A type 1 hexaosylceramide. Finally, the mono-, di-, and triglycosylceramides were characterized as galactosylceramide, glucosylceramide, lactosylceramide, galabiaosylceramide, globotriaosylceramide, and lactotriaosylceramide. Thus, the glycan diversity of human embryonic stem cells, including cell surface immune determinants, is more complex than previously appreciated.
authors: Department of Surgery, Sahlgrenska University Hospital, S-41 345 Göteborg, Sweden.
authors: Angela
authors: A
authors: Barone
authors: Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology, P.O. Box 440, University of Gothenburg, S-405 30 Göteborg, Sweden.
authors: John
authors: J
authors: Benktander
authors: Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology, P.O. Box 440, University of Gothenburg, S-405 30 Göteborg, Sweden.
authors: Jonas
authors: J
authors: Ångström
authors: Cellectis Stem Cells, Cellartis AB, Arvid Wallgrens Backe 20, S-413 46 Göteborg, Sweden.
authors: Anders
authors: A
authors: Aspegren
authors: Cellectis Stem Cells, Cellartis AB, Arvid Wallgrens Backe 20, S-413 46 Göteborg, Sweden.
authors: Petter
authors: P
authors: Björquist
authors: Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology, P.O. Box 440, University of Gothenburg, S-405 30 Göteborg, Sweden. Electronic address: Susann.Teneberg@medkem.gu.se.
authors: Susann
authors: S
authors: Teneberg
authors: Department of Surgery, Sahlgrenska University Hospital, S-41 345 Göteborg, Sweden.
authors: Michael E
authors: ME
authors: Breimer
mesh_terms: D000818:Animals; D002241:Carbohydrates / Q000737:chemistry; D018929:Cell Culture Techniques; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002855:Chromatography, Thin Layer / Q000379:methods; D003470:Culture Media / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology*; D000939:Epitopes / Q000737:chemistry; D005347:Fibroblasts / Q000166:cytology; D006001:Glycoconjugates / Q000737:chemistry; D006017:Glycolipids / Q000737:chemistry; D006028:Glycosphingolipids / Q000737:chemistry*; D006801:Humans; D037102:Lectins / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D051379:Mice; D044968:Regenerative Medicine / Q000379:methods; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
chemical_list: D002241:Carbohydrates; D003470:Culture Media; D000939:Epitopes; D006001:Glycoconjugates; D006017:Glycolipids; D006028:Glycosphingolipids; D037102:Lectins
references: J Am Soc Mass Spectrom. 2002 Jun;13(6):670-9
references: 12056567
references: Methods Enzymol. 1990;193:623-46
references: 2074839
references: Glycoconj J. 2009 Apr;26(3):335-47
references: 18773292
references: Xenotransplantation. 2011 Jul-Aug;18(4):215-28
references: 21848538
references: Science. 1998 Nov 6;282(5391):1145-7
references: 9804556
references: J Mol Cell Biol. 2011 Apr;3(2):99-107
references: 21149348
references: Biochim Biophys Acta. 2008 Mar;1780(3):325-46
references: 17976918
references: Glycoconj J. 2009 Nov;26(8):929-33
references: 18415015
references: Biochem J. 2010 Nov 15;432(1):21-33
references: 20854259
references: Biochem Biophys Res Commun. 1983 Jun 29;113(3):967-74
references: 6307308
references: Chem Phys Lipids. 1988 Nov;49(1-2):31-7
references: 3233709
references: Glycobiology. 2004 Jan;14(1):1-12
references: 14514713
references: J Biochem. 1991 Jul;110(1):120-31
references: 1939018
references: Methods Enzymol. 1987;138:212-20
references: 3600323
references: Stem Cell Res. 2009 Jul;3(1):3-11
references: 19398226
references: Glycobiology. 2010 Sep;20(9):1103-16
references: 20466655
references: Dev Biol. 1987 Jul;122(1):21-34
references: 3297853
references: J Biol Chem. 1983 Apr 10;258(7):4091-7
references: 6833243
references: J Biol Chem. 1982 Jan 25;257(2):906-12
references: 6798034
references: J Biol Chem. 2011 Jun 10;286(23):20345-53
references: 21471226
references: Biochemistry. 1989 Mar 7;28(5):2229-38
references: 2497776
references: Nature. 1981 Jul 9;292(5819):154-6
references: 7242681
references: J Biol Chem. 1989 Jun 5;264(16):9510-9
references: 2722847
references: Proc Natl Acad Sci U S A. 2010 Dec 28;107(52):22564-9
references: 21149694
references: Rapid Commun Mass Spectrom. 2004;18(4):412-20
references: 14966848
references: Proc Natl Acad Sci U S A. 2003 Oct 14;100(21):12045-50
references: 14523234
references: J Biol Chem. 1984 Apr 10;259(7):4672-80
references: 6707026
references: J Biochem. 1990 Nov;108(5):766-77
references: 2081734
references: Stem Cells. 2011 Dec;29(12):1995-2004
references: 21956927
references: J Biol Chem. 2012 Sep 14;287(38):31712-24
references: 22822069
references: Glycobiology. 2007 Aug;17(8):17C-23C
references: 17483136
references: Annu Rev Biochem. 2004;73:491-537
references: 15189151
references: Neurochem Res. 2011 Sep;36(9):1623-35
references: 21161592
references: Annu Rev Cell Dev Biol. 2001;17:435-62
references: 11687496
references: J Biol Chem. 1983 Jul 25;258(14):8934-42
references: 6863318
references: J Biol Chem. 1994 Mar 18;269(11):8554-63
references: 8132581
references: J Biol Chem. 1997 Dec 19;272(51):31979-91
references: 9405390
references: Biochemistry. 1983 May 24;22(11):2676-87
references: 6307345
references: Biol Reprod. 1995 Apr;52(4):903-12
references: 7780013
references: Dev Biol. 1986 Mar;114(1):12-21
references: 2869992
references: Eur J Biochem. 1992 Oct 15;209(2):483-501
references: 1358608
references: Vaccine. 2011 Nov 8;29(48):8802-26
references: 21964054
references: Proc Natl Acad Sci U S A. 1987 Mar;84(5):1369-73
references: 2434954
references: Curr Biol. 1998 Nov 5;8(22):R802-4
references: 9811599
references: Transplantation. 2007 Nov 27;84(10):1348-56
references: 18049121
references: EMBO J. 1983;2(12):2355-61
references: 6141938
references: Eur J Biochem. 1998 Oct 15;257(2):293-8
references: 9826173
references: Glycobiology. 1993 Apr;3(2):97-130
references: 8490246
references: Glycoconj J. 2008 Aug;25(6):561-72
references: 18247115
references: J Anat. 2002 Mar;200(Pt 3):249-58
references: 12033729
references: J Biol Chem. 1998 Nov 6;273(45):29331-40
references: 9792633
references: Nat Med. 2005 Feb;11(2):228-32
references: 15685172