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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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Anion-exchange DEAE elution profiles of PA-glycans.PA-glycans were fractionated according to their sialic acid content as neutral (peak 1), monosialyl (peak 3), and disialyl (peak 4) oligosaccharide fractions. Peaks 2 and 5 represent fractions containing no detectable PA-oligosaccharides.
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pone-0111064-g004: Anion-exchange DEAE elution profiles of PA-glycans.PA-glycans were fractionated according to their sialic acid content as neutral (peak 1), monosialyl (peak 3), and disialyl (peak 4) oligosaccharide fractions. Peaks 2 and 5 represent fractions containing no detectable PA-oligosaccharides.

Mentions: N-glycans extracted from undifferentiated iPSCs (959A2-1: 26 mg, 959C1-1: 11 mg and 956F-1: 10 mg of protein), iPSC-CM (959A2-1 CM: 15 mg, 959C1-1 CM: 12 mg and 956F-1 CM: 5.5 mg of protein), and the B6 heart muscle (82 mg of protein) were separated on a diethylaminoethyl (DEAE) column into five peaks, based on increasing acidity. Peak 1 represented a neutral (N) fraction, peak 3 a monosialyl (M) fraction, and peak 4 a disialyl (D) fraction. Glycan fractions in each of these peaks were as follows: iPSCs yielded 97% N, 0.5% M, 2.6% D (959A2-1), 98% N, 0.7% M, 1.1% D (959C1-1) and 96% N, 1.1% M, 3.1% D (956F-1) peak areas, iPSC-CMs yielded 89% N, 6.4% M, 4.4% D (959A2-1 CM), 79% N, 16% M, 4.8% D (959C1-1 CM) and 82% N, 10% M, 7.9% D (956F-1 CM) and Heart yielded 55% N, 19% M, and 25% D (Figure 4).


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)

Anion-exchange DEAE elution profiles of PA-glycans.PA-glycans were fractionated according to their sialic acid content as neutral (peak 1), monosialyl (peak 3), and disialyl (peak 4) oligosaccharide fractions. Peaks 2 and 5 represent fractions containing no detectable PA-oligosaccharides.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0111064-g004: Anion-exchange DEAE elution profiles of PA-glycans.PA-glycans were fractionated according to their sialic acid content as neutral (peak 1), monosialyl (peak 3), and disialyl (peak 4) oligosaccharide fractions. Peaks 2 and 5 represent fractions containing no detectable PA-oligosaccharides.
Mentions: N-glycans extracted from undifferentiated iPSCs (959A2-1: 26 mg, 959C1-1: 11 mg and 956F-1: 10 mg of protein), iPSC-CM (959A2-1 CM: 15 mg, 959C1-1 CM: 12 mg and 956F-1 CM: 5.5 mg of protein), and the B6 heart muscle (82 mg of protein) were separated on a diethylaminoethyl (DEAE) column into five peaks, based on increasing acidity. Peak 1 represented a neutral (N) fraction, peak 3 a monosialyl (M) fraction, and peak 4 a disialyl (D) fraction. Glycan fractions in each of these peaks were as follows: iPSCs yielded 97% N, 0.5% M, 2.6% D (959A2-1), 98% N, 0.7% M, 1.1% D (959C1-1) and 96% N, 1.1% M, 3.1% D (956F-1) peak areas, iPSC-CMs yielded 89% N, 6.4% M, 4.4% D (959A2-1 CM), 79% N, 16% M, 4.8% D (959C1-1 CM) and 82% N, 10% M, 7.9% D (956F-1 CM) and Heart yielded 55% N, 19% M, and 25% D (Figure 4).

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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