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.
Highlights
In vitro generation of cardiac myocytes by reprogramming is a promising technology in developing cell-transplant therapy for advanced cardiac failure [1] and drug discovery for a variety of cardiac diseases [2]
Purified cardiomyocytes derived from induced pluripotent stem cells (iPSCs) Cardiomyogenic differentiation was induced in murine iPSCs by using a slightly modified culture protocol (Figure 1a)
The iPSC-derived cardiomyocytes (iPSC-CM) were functional with Ca2+ transient measurement (Figure 3a, b) and their beating rates were increased by the administration of isoproterenol (Figure 3c), meaning they had b-adrenergic receptors
Summary
In vitro generation of cardiac myocytes by reprogramming is a promising technology in developing cell-transplant therapy for advanced cardiac failure [1] and drug discovery for a variety of cardiac diseases [2]. For both purposes, induced pluripotent stem cells (iPSCs) are most useful, since generation and cardiomyogenic differentiation of iPSCs has been standardized in human and a number of animals [3,4]. Derivatives of iPSCs have been developed to the pre-clinical stage for cell transplantation therapy [5], while cardiac myocytes generated from patient-specific iPSCs have been studied to explore pathologic mechanisms and guide drug discovery [6,7]. We analyzed N-glycan expression in undifferentiated iPSCs, iPSC-CMs, and adult murine myocardium by HPLC, to identify potential indicators of the maturity of differentiating cardiomyocytes from iPS cells in vitro
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