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N-glycans: phenotypic homology and structural differences between myocardial cells and induced pluripotent stem cell-derived cardiomyocytes.

Kawamura T, Miyagawa S, Fukushima S, Yoshida A, Kashiyama N, Kawamura A, Ito E, Saito A, Maeda A, Eguchi H, Toda K, Lee JK, Miyagawa S, Sawa Y - PLoS ONE (2014)

Bottom Line: The quantity of high-mannose type (immature) N-glycans on the iPSCs decreased with cardiomyogenic differentiation, but did not reach the low levels observed in the heart.We observed a similar reduction in neutral N-glycans and an increase in fucosylated or sialyl N-glycans.The expression of N-glycan on murine iPSCs changed toward the myocardial phenotype during cardiomyogenic differentiation, leaving the structural differences of NeuGc content or Galα1-6 Gal structures.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiovascular Surgery, Osaka University Graduate School of Medicine, Suita, Osaka, Japan.

ABSTRACT
Cell surface glycans vary widely, depending on cell properties. We hypothesized that glycan expression on induced pluripotent stem cells (iPSCs) might change during cardiomyogenic differentiation toward the myocardial phenotype. N-glycans were isolated from iPSCs, iPSC-derived cardiomyocytes (iPSC-CM), and original C57BL/6 mouse myocardium (Heart). Their structures were analyzed by a mapping technique based on HPLC elution times and MALDI-TOF/MS spectra. Sixty-eight different N-glycans were isolated; the structures of 60 of these N-glycans were identified. The quantity of high-mannose type (immature) N-glycans on the iPSCs decreased with cardiomyogenic differentiation, but did not reach the low levels observed in the heart. We observed a similar reduction in neutral N-glycans and an increase in fucosylated or sialyl N-glycans. Some structural differences were detected between iPSC-CM and Heart. No N-glycolyl neuraminic acid (NeuGc) structures were detected in iPSC-CM, whereas the heart contained numerous NeuGc structures, corresponding to the expression of cytidine monophosphate-N-acetylneuraminic acid hydroxylase. Furthermore, several glycans containing Galα1-6 Gal, rarely identified in the other cells, were detected in the iPSC-CM. The expression of N-glycan on murine iPSCs changed toward the myocardial phenotype during cardiomyogenic differentiation, leaving the structural differences of NeuGc content or Galα1-6 Gal structures. Further studies will be warranted to reveal the meaning of the difference of N-glycans between the iPSC-CM and the myocardium.

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Structures of neutral, monosialyl, and disialyl PA-oligosaccharides in iPSCs, iPSC-CM, and heart cells.Glucose units (GU) were calculated from the peak elution times for the ODS column in Figure 5, 6 and 7, and the amide column (data not shown). Average mass (Mass) calculated from the mlz values of [M+Na]+ or [M+H]+ ion for neutral, [M-H]− ion for monosialyl, and [M-H]− & [M+Na-2H]− ions for disialyl PA-oligosaccharides.
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pone-0111064-g008: Structures of neutral, monosialyl, and disialyl PA-oligosaccharides in iPSCs, iPSC-CM, and heart cells.Glucose units (GU) were calculated from the peak elution times for the ODS column in Figure 5, 6 and 7, and the amide column (data not shown). Average mass (Mass) calculated from the mlz values of [M+Na]+ or [M+H]+ ion for neutral, [M-H]− ion for monosialyl, and [M-H]− & [M+Na-2H]− ions for disialyl PA-oligosaccharides.

Mentions: N-glycans N9-2, M8, M12, M17, and M23 were trimmed by α-galactosidase but not by β-galactosidase or N-acetylglucosamidase. Their structures fit GALAXY references H5.12, 1A1-200.4, 1A3-200.4, 1A1-210.4, and 1A3-210.4, respectively. The galactosyl structures were then identified as Galα1-6Gal, because of the α-galactosidase-driven MS shifts. The structure of the M13 was identified by the coincidence with a GALAXY reference 1A2-H5.12 after being trimmed by α-L-fucosidase. The other N-glycans M11-2, M15, M18, M19, M20-2, M21, D8 and D9 were not identified in this study because they did not correspond to GALAXY references even after α-galactosidase digestions. They are described in Figure 8 and Table S1-S5 with their proposed formulas based on MALDI-TOF/MS data.


N-glycans: phenotypic homology and structural differences between myocardial cells and induced pluripotent stem cell-derived cardiomyocytes.

Kawamura T, Miyagawa S, Fukushima S, Yoshida A, Kashiyama N, Kawamura A, Ito E, Saito A, Maeda A, Eguchi H, Toda K, Lee JK, Miyagawa S, Sawa Y - PLoS ONE (2014)

Structures of neutral, monosialyl, and disialyl PA-oligosaccharides in iPSCs, iPSC-CM, and heart cells.Glucose units (GU) were calculated from the peak elution times for the ODS column in Figure 5, 6 and 7, and the amide column (data not shown). Average mass (Mass) calculated from the mlz values of [M+Na]+ or [M+H]+ ion for neutral, [M-H]− ion for monosialyl, and [M-H]− & [M+Na-2H]− ions for disialyl PA-oligosaccharides.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4214687&req=5

pone-0111064-g008: Structures of neutral, monosialyl, and disialyl PA-oligosaccharides in iPSCs, iPSC-CM, and heart cells.Glucose units (GU) were calculated from the peak elution times for the ODS column in Figure 5, 6 and 7, and the amide column (data not shown). Average mass (Mass) calculated from the mlz values of [M+Na]+ or [M+H]+ ion for neutral, [M-H]− ion for monosialyl, and [M-H]− & [M+Na-2H]− ions for disialyl PA-oligosaccharides.
Mentions: N-glycans N9-2, M8, M12, M17, and M23 were trimmed by α-galactosidase but not by β-galactosidase or N-acetylglucosamidase. Their structures fit GALAXY references H5.12, 1A1-200.4, 1A3-200.4, 1A1-210.4, and 1A3-210.4, respectively. The galactosyl structures were then identified as Galα1-6Gal, because of the α-galactosidase-driven MS shifts. The structure of the M13 was identified by the coincidence with a GALAXY reference 1A2-H5.12 after being trimmed by α-L-fucosidase. The other N-glycans M11-2, M15, M18, M19, M20-2, M21, D8 and D9 were not identified in this study because they did not correspond to GALAXY references even after α-galactosidase digestions. They are described in Figure 8 and Table S1-S5 with their proposed formulas based on MALDI-TOF/MS data.

Bottom Line: The quantity of high-mannose type (immature) N-glycans on the iPSCs decreased with cardiomyogenic differentiation, but did not reach the low levels observed in the heart.We observed a similar reduction in neutral N-glycans and an increase in fucosylated or sialyl N-glycans.The expression of N-glycan on murine iPSCs changed toward the myocardial phenotype during cardiomyogenic differentiation, leaving the structural differences of NeuGc content or Galα1-6 Gal structures.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiovascular Surgery, Osaka University Graduate School of Medicine, Suita, Osaka, Japan.

ABSTRACT
Cell surface glycans vary widely, depending on cell properties. We hypothesized that glycan expression on induced pluripotent stem cells (iPSCs) might change during cardiomyogenic differentiation toward the myocardial phenotype. N-glycans were isolated from iPSCs, iPSC-derived cardiomyocytes (iPSC-CM), and original C57BL/6 mouse myocardium (Heart). Their structures were analyzed by a mapping technique based on HPLC elution times and MALDI-TOF/MS spectra. Sixty-eight different N-glycans were isolated; the structures of 60 of these N-glycans were identified. The quantity of high-mannose type (immature) N-glycans on the iPSCs decreased with cardiomyogenic differentiation, but did not reach the low levels observed in the heart. We observed a similar reduction in neutral N-glycans and an increase in fucosylated or sialyl N-glycans. Some structural differences were detected between iPSC-CM and Heart. No N-glycolyl neuraminic acid (NeuGc) structures were detected in iPSC-CM, whereas the heart contained numerous NeuGc structures, corresponding to the expression of cytidine monophosphate-N-acetylneuraminic acid hydroxylase. Furthermore, several glycans containing Galα1-6 Gal, rarely identified in the other cells, were detected in the iPSC-CM. The expression of N-glycan on murine iPSCs changed toward the myocardial phenotype during cardiomyogenic differentiation, leaving the structural differences of NeuGc content or Galα1-6 Gal structures. Further studies will be warranted to reveal the meaning of the difference of N-glycans between the iPSC-CM and the myocardium.

Show MeSH