Structural Complexity of Non-acid Glycosphingolipids in Human Embryonic Stem Cells Grown under Feeder-free Conditions

Abstract

Background: Carbohydrate epitopes are often used as markers for characterization of human embryonic stem cells (hESC). Results: Several glycosphingolipids not previously found in hESC were characterized. Conclusion: The glycosylation of hESC is more complex than previously thought. Significance: These findings will help to understand the immunogenicity of hESC and might impact future applications in regenerative medicine.

A Cytotoxic Antibody Recognizing Lacto-N-fucopentaose I (LNFP I) on Human Induced Pluripotent Stem (hiPS) Cells

“…A switch of the core structures of glycans from the globo-and lacto-series (type 1 structure) to the ganglio-series during hES cell differentiation has been reported (27). Similarly, the lacto-N-fucopentaose I structure has been shown to be a characteristic feature of undifferentiated hiPS/ES cells by extensive structural studies on hiPS/ES cell glycolipids (28,29), and this was supported by lectin array (30) and glycan array (31) studies. In addition, R-17F binds dose-dependently to chemically synthesized biotinylated LNFP I on avidin-coated plates, so we have developed an ELISA system to quantitate the binding activity of every lot of R-17F antibodies (data not shown).…”

Glycosphingolipids of porcine, bovine, and equine pericardia as potential immune targets in bioprosthetic heart valve grafts

“…There was no 0,2 A ion at m/z 630, or 0,2 A‐H 2 O ion at m/z 612, demonstrating 4‐substitution of the Gal of the lactose unit at the reducing end and thus identifying Fuc‐gangliotetra, and no binding of anti‐H type 1 antibodies to the non‐acid fraction of porcine pericards was obtained (see below). However, on graphite columns, the H type 1 and the H type 2 pentasaccharides co‐elute, and here, the [M‐H + ] − at m/z 852 eluted at 22.5 minutes, that is, earlier than the H type 2 pentasaccharide. Thus, the molecular ion at m/z 852 eluting at 22.5 minutes was tentatively identified as a Fuc‐ganglio pentasaccharide.…”

“…Endoglycoceramidase II from Rhodococcus spp. (Takara Bio Europe S.A., Gennevilliers, France) was used for hydrolysis of the total non‐acid glycosphingolipid fractions to obtain free oligosaccharides . The glycosphingolipid‐derived oligosaccharides were analyzed by liquid chromatography electron spray ionization mass spectrometry (LC‐ESI/MS) as described …”

Glycolipid dynamics in generation and differentiation of induced pluripotent stem cells

“…The neutral GSLs were detected as Hex-Cer, Hex-Hex-Cer, and Hex-Hex-Hex-Cer, HexNAc-Hex-Hex-Hex-Cer by MS/MS analysis. Though the neutral GSLs have isomers depending on the variety of monosaccharide, their structures were deduced to be GlcCer, LacCer, Gb3Cer, Gb4Cer because they are major GSLs and detected even in hESCs 18 . However, since GalCer and galabiose as isomers of GlcCer and LacCer have been detected in hESCs 18 , existence of other isomers in hiPSCs could not be excluded.…”

“…Though those sequences have isomers, the structures were deduced from the MS/MS analysis and immunocytochemical analysis described later. They were major GSLs synthesized by biosynthetic pathways expressed in hESCs 16 17 18 19 . This is the first time that the structures for Gb5Cer, sialyl-Gb5Cer, fucosyl-Gb5Cer, and IV fucosyl-(n)Lc4Cer were identified in undifferentiated hiPSCs.…”