Glycome Diagnosis of Human Induced Pluripotent Stem Cells Using Lectin Microarray

Induced pluripotent stem cells (iPSCs) can now be produced from various somatic cell (SC) lines by ectopic expression of the four transcription factors. Although the procedure has been demonstrated to induce global change in gene and microRNA expressions and even epigenetic modification, it remains largely unknown how this transcription factor-induced reprogramming affects the total glycan repertoire expressed on the cells. Here we performed a comprehensive glycan analysis using 114 types of human iPSCs generated from five different SCs and compared their glycomes with those of human embryonic stem cells (ESCs; nine cell types) using a high density lectin microarray. In unsupervised cluster analysis of the results obtained by lectin microarray, both undifferentiated iPSCs and ESCs were clustered as one large group. However, they were clearly separated from the group of differentiated SCs, whereas all of the four SCs had apparently distinct glycome profiles from one another, demonstrating that SCs with originally distinct glycan profiles have acquired those similar to ESCs upon induction of pluripotency. Thirty-eight lectins discriminating between SCs and iPSCs/ESCs were statistically selected, and characteristic features of the pluripotent state were then obtained at the level of the cellular glycome. The expression profiles of relevant glycosyltransferase genes agreed well with the results obtained by lectin microarray. Among the 38 lectins, rBC2LCN was found to detect only undifferentiated iPSCs/ESCs and not differentiated SCs. Hence, the high density lectin microarray has proved to be valid for not only comprehensive analysis of glycans but also diagnosis of stem cells under the concept of the cellular glycome. Induced pluripotent stem cells (iPSCs) can now be produced from various somatic cell (SC) lines by ectopic expression of the four transcription factors. Although the procedure has been demonstrated to induce global change in gene and microRNA expressions and even epigenetic modification, it remains largely unknown how this transcription factor-induced reprogramming affects the total glycan repertoire expressed on the cells. Here we performed a comprehensive glycan analysis using 114 types of human iPSCs generated from five different SCs and compared their glycomes with those of human embryonic stem cells (ESCs; nine cell types) using a high density lectin microarray. In unsupervised cluster analysis of the results obtained by lectin microarray, both undifferentiated iPSCs and ESCs were clustered as one large group. However, they were clearly separated from the group of differentiated SCs, whereas all of the four SCs had apparently distinct glycome profiles from one another, demonstrating that SCs with originally distinct glycan profiles have acquired those similar to ESCs upon induction of pluripotency. Thirty-eight lectins discriminating between SCs and iPSCs/ESCs were statistically selected, and characteristic features of the pluripotent state were then obtained at the level of the cellular glycome. The expression profiles of relevant glycosyltransferase genes agreed well with the results obtained by lectin microarray. Among the 38 lectins, rBC2LCN was found to detect only undifferentiated iPSCs/ESCs and not differentiated SCs. Hence, the high density lectin microarray has proved to be valid for not only comprehensive analysis of glycans but also diagnosis of stem cells under the concept of the cellular glycome. IntroductionIncreasing attention has been paid to iPSCs 2The abbreviations used are: iPSCinduced pluripotent stem cellAMamniotic mesodermalESCembryonic stem cellMEFmouse embryonic fibroblastPAEplacental artery endothelialSCsomatic cellUtEuterine endometriumrMOArecombinant MOAFWERfamilywise error rate. and ESCs in their pluripotency and medical applications (1Takahashi K. Tanabe K. Ohnuki M. Narita M. Ichisaka T. Tomoda K. Yamanaka S. Cell. 2007; 131: 861-872Abstract Full Text Full Text PDF PubMed Scopus (14653) Google Scholar, 2Yu J. Vodyanik M.A. Smuga-Otto K. Antosiewicz-Bourget J. Frane J.L. Tian S. Nie J. Jonsdottir G.A. Ruotti V. Stewart R. Slukvin I.I. Thomson J.A. Science. 2007; 318: 1917-1920Crossref PubMed Scopus (8043) Google Scholar). However, establishment of a robust evaluation system of their properties, including differentiation propensity and risk of possible contamination of xenoantigens and even potential of tumorigenesis, has been hampered by the lack of comprehensive methodology directly applicable to target stem cells, although this is an emerging issue essential for the safe use of iPSCs in regenerative medicine. From many aspects, cell surface glycans are considered to be ideal targets for analyzing or identifying the phenotype of each cell in a direct manner by the following reasons (3Gagneux P. Varki A. Glycobiology. 1999; 9: 747-755Crossref PubMed Scopus (421) Google Scholar, 4Varki A. Glycobiology. 1993; 3: 97-130Crossref PubMed Scopus (4963) Google Scholar). (a) Glycans are located at the outermost cell surface. (b) The total repertoire of cell surface glycans varies at every level of biological organization (i.e. species, tissues, cell types, and molecules). (c) Global alterations of the cellular glycome also occur during development, cellular activation, differentiation, malignant transformation, and inflammation. The cell surface glycans are therefore referred to as the “cell signature” that closely reflects cellular backgrounds and conditions, probably because they are actually functioning as cell-to-cell mediators in extensive biological phenomena. This fundamental nature of glycans should be understood with the fact that they are not encoded directly in the genome but are generated by a complex system of a number of glycosidases and glycosyltransferases, whose expressions and activities are significantly affected by both intracellular and extracellular environmental changes. Indeed, cell surface molecules, such as stage-specific embryonic antigens (SSEA1 and -3/4) (5Muramatsu T. Muramatsu H. Glycoconj. J. 2004; 21: 41-45Crossref PubMed Scopus (116) Google Scholar) and tumor rejection antigens (Tra-1-60 and Tra-1-81) (6Schopperle W.M. DeWolf W.C. Stem Cells. 2007; 25: 723-730Crossref PubMed Scopus (106) Google Scholar, 7Natunen S. Satomaa T. Pitkanen V. Salo H. Mikkola M. Natunen J. Otonkoski T. Valmu L. Glycobiology. 2011; (in press)PubMed Google Scholar, 8Lanctot P.M. Gage F.H. Varki A.P. Curr. Opin. Chem. Biol. 2007; 11: 373-380Crossref PubMed Scopus (130) Google Scholar) are glycobiomarkers widely used to evaluate pluripotency. Notably, however, these “representative” glycomarkers have been identified following rather fortuitous development of their specific antibodies, because most carbohydrate structures are poorly antigenic between mammals. In this context, a systematic search is necessary to draw a whole picture of the stem cell glycome and harness its effect on stem cell biology (8Lanctot P.M. Gage F.H. Varki A.P. Curr. Opin. Chem. Biol. 2007; 11: 373-380Crossref PubMed Scopus (130) Google Scholar, 9Liang Y.J. Kuo H.H. Lin C.H. Chen Y.Y. Yang B.C. Cheng Y.Y. Yu A.L. Khoo K.H. Yu J. Proc. Natl. Acad. Sci. U.S.A. 2010; 107: 22564-22569Crossref PubMed Scopus (86) Google Scholar). For instance, the growth and directed differentiation of stem cells to specific progeny lineages in cell culture remain problematic. Understanding how stem cells communicate with one another and feeder cells through cell surface glycans may lead to rational design of specific culture systems. However, the glycome is a quite difficult target to predict solely based on any genomic data base because the biosynthetic process of the glycan moieties of glycoproteins is not template-driven and is subject to multiple sequential and competitive enzymatic pathways. In this sense, a rapid and sensitive system enabling direct monitoring of cell surface glycans is essential.Several methods have been developed for glycan analysis based on physicochemical principles, such as liquid chromatography and mass spectrometry (10Hirabayashi J. J. Biochem. 2008; 144: 139-147Crossref PubMed Scopus (121) Google Scholar, 11Wearne K.A. Winter H.C. Goldstein I.J. Glycoconj. J. 2008; 25: 121-136Crossref PubMed Scopus (15) Google Scholar, 12Wearne K.A. Winter H.C. O'Shea K. Goldstein I.J. Glycobiology. 2006; 16: 981-990Crossref PubMed Scopus (57) Google Scholar). Lectin microarray is an alternative technology for structural glycomics, where a panel of lectins with various glycan-binding specificities is printed on a microarray, providing a versatile platform for rapid and high throughput analysis of glycan structures without liberation of glycans (13Pilobello K.T. Krishnamoorthy L. Slawek D. Mahal L.K. Chembiochem. 2005; 6: 985-989Crossref PubMed Scopus (244) Google Scholar, 14Kuno A. Uchiyama N. Koseki-Kuno S. Ebe Y. Takashima S. Yamada M. Hirabayashi J. Nat. Methods. 2005; 2: 851-856Crossref PubMed Scopus (433) Google Scholar). Lectins are a class of decoder molecules of cell surface glycans distributed throughout organisms, which mediate various functions through specific glycan recognition. Analytical protocols using lectin microarray have been developed for various sample types: free oligosaccharides (14Kuno A. Uchiyama N. Koseki-Kuno S. Ebe Y. Takashima S. Yamada M. Hirabayashi J. Nat. Methods. 2005; 2: 851-856Crossref PubMed Scopus (433) Google Scholar, 15Uchiyama N. Kuno A. Tateno H. Kubo Y. Mizuno M. Noguchi M. Hirabayashi J. Proteomics. 2008; 8: 3042-3050Crossref PubMed Scopus (51) Google Scholar), tissue sections (16Matsuda A. Kuno A. Ishida H. Kawamoto T. Shoda J. Hirabayashi J. Biochem. Biophys. Res. Commun. 2008; 370: 259-263Crossref PubMed Scopus (58) Google Scholar), cell membrane hydrophobic fractions (17Ebe Y. Kuno A. Uchiyama N. Koseki-Kuno S. Yamada M. Sato T. Narimatsu H. Hirabayashi J. J. Biochem. 2006; 139: 323-327Crossref PubMed Scopus (57) Google Scholar, 18Pilobello K.T. Slawek D.E. Mahal L.K. Proc. Natl. Acad. Sci. U.S.A. 2007; 104: 11534-11539Crossref PubMed Scopus (175) Google Scholar), and even whole cells (19Hsu K.L. Pilobello K.T. Mahal L.K. Nat. Chem. Biol. 2006; 2: 153-157Crossref PubMed Scopus (208) Google Scholar, 20Tateno H. Uchiyama N. Kuno A. Togayachi A. Sato T. Narimatsu H. Hirabayashi J. Glycobiology. 2007; 17: 1138-1146Crossref PubMed Scopus (136) Google Scholar). This technology has just begun to be applied to a wide variety of biological researches, including virus profiling (21Krishnamoorthy L. Bess Jr., J.W. Preston A.B. Nagashima K. Mahal L.K. Nat. Chem. Biol. 2009; 5: 244-250Crossref PubMed Scopus (133) Google Scholar) and cell profiling (17Ebe Y. Kuno A. Uchiyama N. Koseki-Kuno S. Yamada M. Sato T. Narimatsu H. Hirabayashi J. J. Biochem. 2006; 139: 323-327Crossref PubMed Scopus (57) Google Scholar, 20Tateno H. Uchiyama N. Kuno A. Togayachi A. Sato T. Narimatsu H. Hirabayashi J. Glycobiology. 2007; 17: 1138-1146Crossref PubMed Scopus (136) Google Scholar), and development of cancer glycobiomarkers (22Kuno A. Kato Y. Matsuda A. Kaneko M.K. Ito H. Amano K. Chiba Y. Narimatsu H. Hirabayashi J. Mol. Cell Proteomics. 2009; 8: 99-108Abstract Full Text Full Text PDF PubMed Scopus (100) Google Scholar, 23Narimatsu H. Sawaki H. Kuno A. Kaji H. Ito H. Ikehara Y. FEBS J. 2010; 277: 95-105Crossref PubMed Scopus (146) Google Scholar, 24Matsuda A. Kuno A. Kawamoto T. Matsuzaki H. Irimura T. Ikehara Y. Zen Y. Nakanuma Y. M. N. Shoda J. Hirabayashi J. Narimatsu H. 2010; PubMed Scopus Google Scholar). For cell of in hydrophobic fractions are for each analysis H. Kuno A. Y. Hirabayashi J. 2010; PubMed Scopus Google Scholar, A. Y. M. Y. Yamada M. A. Hirabayashi J. J. 2008; Google Scholar). and were to the of the data H. Kuno A. Y. Hirabayashi J. 2010; PubMed Scopus Google Scholar, A. Y. M. Y. Yamada M. A. Hirabayashi J. J. 2008; Google Scholar). we have demonstrated that lectin microarray is also applicable to stem cells M. M. Y. Kuno A. T. Yamada M. H. Y. S. Narimatsu H. Hirabayashi J. A. Cells. 2011; 16: PubMed Scopus Google Scholar, S. Y. Ito Y. Tateno H. M. H. K. Y. Y. H. N. Y. A. Hirabayashi J. K. M. The on Scholar), although we have to a as to how the cellular glycome upon induction of pluripotency. of this technology to the of stem cells has not been we developed an platform of high density lectin microarray with the number of lectins to the glycome for of various stem cell systematic of the cellular glycome was then performed cell types in including iPSCs cell types) and ESCs cell this comprehensive we obtained that all of the four SCs with originally distinct glycan profiles have acquired those similar to ESCs upon induction of pluripotency. also found structural features to iPSCs and which well to the results of gene expression analysis of we the of lectin microarray in the stem cell diagnosis of multiple including between undifferentiated and differentiated cells as well as of the contamination of the the developed high density lectin microarray, we performed a systematic analysis of cell surface glycans of a large of human iPSCs cell types) and ESCs cell a a for a rational stem cell evaluation system was which can both the state of and of a comprehensive glycome analysis iPSCs and ESCs has been are at in using a lectin microarray. glycan of each cell is obtained using a number of cells and the is widely applicable to stem cells. The evaluation system of the by a a number of lectins, which are on the developed also be of stem cells can be (i.e. with the in this evaluation methods and the differentiation propensity of stem cells also be on the lectin and the expression profiles of glycosyltransferases, we that the expression of and whereas that of and upon the induction of pluripotency. are with the that relevant glycans (i.e. the expression of and with an and in human are that in differentiated Y.J. Kuo H.H. Lin C.H. Chen Y.Y. Yang B.C. Cheng Y.Y. Yu A.L. Khoo K.H. Yu J. Proc. Natl. Acad. Sci. U.S.A. 2010; 107: 22564-22569Crossref PubMed Scopus (86) Google Scholar, T. A. Mikkola M. M. M. T. A. J. J. Natunen J. T. Otonkoski T. J. J. Cell Biol. 2009; PubMed Scopus Google Scholar). the expression of and which are by the of and is closely to the of the well pluripotency and a this rBC2LCN was as the lectin to evaluate pluripotency the is a lectin identified from a M. M. A. A. M. 2010; Full Text Full Text PDF PubMed Scopus (58) Google Scholar). microarray analysis that rBC2LCN to such as and which the structural to the pluripotency and as This is with the M. M. A. A. M. 2010; Full Text Full Text PDF PubMed Scopus (58) Google Scholar) in which the glycan-binding of in using glycan microarray and also demonstrated that this lectin also to which was as a pluripotency Y.J. Kuo H.H. Lin C.H. Chen Y.Y. Yang B.C. Cheng Y.Y. Yu A.L. Khoo K.H. Yu J. Proc. Natl. Acad. Sci. U.S.A. 2010; 107: 22564-22569Crossref PubMed Scopus (86) Google Scholar, M. M. A. A. M. 2010; Full Text Full Text PDF PubMed Scopus (58) Google Scholar). results the of how this lectin be used as the to pluripotency. rBC2LCN be used to glycoproteins and all whereas and target From a rBC2LCN is because it can be produced in large by the expression system this lectin be a versatile to evaluate has also been to be in cell S. R. S. R. S. J. Biol. Chem. Full Text PDF PubMed Google Scholar). in iPSCs/ESCs has been to be in many types of human and its high expression with tumor and Y. J. Biol. Chem. 2011; Full Text Full Text PDF PubMed Scopus (121) Google Scholar). the glycan alterations upon induction of pluripotency in this are apparently similar to those during malignant transformation, as was Y.J. Kuo H.H. Lin C.H. Chen Y.Y. Yang B.C. Cheng Y.Y. Yu A.L. Khoo K.H. Yu J. Proc. Natl. Acad. Sci. U.S.A. 2010; 107: 22564-22569Crossref PubMed Scopus (86) Google Scholar). Although the for this remains to be the characteristic glycan should be to the of cell and to both cancer cells and pluripotent stem are located at the outermost cell where various on the of cell-to-cell and lectins, molecules of should in the by In this context, between cell surface glycans and lectins are considered to be essential for the of and differentiation of iPSCs/ESCs L. Nat. Methods. 2010; PubMed Scopus Google Scholar). Indeed, were to and pluripotency of embryonic stem cells N. K. K. K. A. H. T. Y. M. K. S. K. S. J. Biol. Chem. 2008; Full Text Full Text PDF PubMed Scopus Google Scholar). on and were demonstrated to direct differentiation of ESCs and human iPSCs N. T. K. M. Y. H. N. H. A. S. Biochem. Biophys. Res. Commun. 2010; PubMed Scopus Google Scholar). cell surface glycans were to the culture of pluripotent stem cells L. Nat. Methods. 2010; PubMed Scopus Google Scholar). global analysis of the cellular glycomes of iPSCs and ESCs performed in this be necessary to the to the functions and applications of the stem cell rational design of the and culture to the of ESCs and iPSCs L. Nat. Methods. 2010; PubMed Scopus Google Scholar). the results obtained in this also be applied to of the the of necessary and of specific cells of undifferentiated In this stem cell with the of a lectin microarray is a issue in of regenerative in the IntroductionIncreasing attention has been paid to iPSCs 2The abbreviations used are: iPSCinduced pluripotent stem cellAMamniotic mesodermalESCembryonic stem cellMEFmouse embryonic fibroblastPAEplacental artery endothelialSCsomatic cellUtEuterine endometriumrMOArecombinant MOAFWERfamilywise error rate. and ESCs in their pluripotency and medical applications (1Takahashi K. Tanabe K. Ohnuki M. Narita M. Ichisaka T. Tomoda K. Yamanaka S. Cell. 2007; 131: 861-872Abstract Full Text Full Text PDF PubMed Scopus (14653) Google Scholar, 2Yu J. Vodyanik M.A. Smuga-Otto K. Antosiewicz-Bourget J. Frane J.L. Tian S. Nie J. Jonsdottir G.A. Ruotti V. Stewart R. Slukvin I.I. Thomson J.A. Science. 2007; 318: 1917-1920Crossref PubMed Scopus (8043) Google Scholar). However, establishment of a robust evaluation system of their properties, including differentiation propensity and risk of possible contamination of xenoantigens and even potential of tumorigenesis, has been hampered by the lack of comprehensive methodology directly applicable to target stem cells, although this is an emerging issue essential for the safe use of iPSCs in regenerative medicine. From many aspects, cell surface glycans are considered to be ideal targets for analyzing or identifying the phenotype of each cell in a direct manner by the following reasons (3Gagneux P. Varki A. Glycobiology. 1999; 9: 747-755Crossref PubMed Scopus (421) Google Scholar, 4Varki A. Glycobiology. 1993; 3: 97-130Crossref PubMed Scopus (4963) Google Scholar). (a) Glycans are located at the outermost cell surface. (b) The total repertoire of cell surface glycans varies at every level of biological organization (i.e. species, tissues, cell types, and molecules). (c) Global alterations of the cellular glycome also occur during development, cellular activation, differentiation, malignant transformation, and inflammation. The cell surface glycans are therefore referred to as the “cell signature” that closely reflects cellular backgrounds and conditions, probably because they are actually functioning as cell-to-cell mediators in extensive biological phenomena. This fundamental nature of glycans should be understood with the fact that they are not encoded directly in the genome but are generated by a complex system of a number of glycosidases and glycosyltransferases, whose expressions and activities are significantly affected by both intracellular and extracellular environmental changes. Indeed, cell surface molecules, such as stage-specific embryonic antigens (SSEA1 and -3/4) (5Muramatsu T. Muramatsu H. Glycoconj. J. 2004; 21: 41-45Crossref PubMed Scopus (116) Google Scholar) and tumor rejection antigens (Tra-1-60 and Tra-1-81) (6Schopperle W.M. DeWolf W.C. Stem Cells. 2007; 25: 723-730Crossref PubMed Scopus (106) Google Scholar, 7Natunen S. Satomaa T. Pitkanen V. Salo H. Mikkola M. Natunen J. Otonkoski T. Valmu L. Glycobiology. 2011; (in press)PubMed Google Scholar, 8Lanctot P.M. Gage F.H. Varki A.P. Curr. Opin. Chem. Biol. 2007; 11: 373-380Crossref PubMed Scopus (130) Google Scholar) are glycobiomarkers widely used to evaluate pluripotency. Notably, however, these “representative” glycomarkers have been identified following rather fortuitous development of their specific antibodies, because most carbohydrate structures are poorly antigenic between mammals. In this context, a systematic search is necessary to draw a whole picture of the stem cell glycome and harness its effect on stem cell biology (8Lanctot P.M. Gage F.H. Varki A.P. Curr. Opin. Chem. Biol. 2007; 11: 373-380Crossref PubMed Scopus (130) Google Scholar, 9Liang Y.J. Kuo H.H. Lin C.H. Chen Y.Y. Yang B.C. Cheng Y.Y. Yu A.L. Khoo K.H. Yu J. Proc. Natl. Acad. Sci. U.S.A. 2010; 107: 22564-22569Crossref PubMed Scopus (86) Google Scholar). For instance, the growth and directed differentiation of stem cells to specific progeny lineages in cell culture remain problematic. Understanding how stem cells communicate with one another and feeder cells through cell surface glycans may lead to rational design of specific culture systems. However, the glycome is a quite difficult target to predict solely based on any genomic data base because the biosynthetic process of the glycan moieties of glycoproteins is not template-driven and is subject to multiple sequential and competitive enzymatic pathways. In this sense, a rapid and sensitive system enabling direct monitoring of cell surface glycans is essential.Several methods have been developed for glycan analysis based on physicochemical principles, such as liquid chromatography and mass spectrometry (10Hirabayashi J. J. Biochem. 2008; 144: 139-147Crossref PubMed Scopus (121) Google Scholar, 11Wearne K.A. Winter H.C. Goldstein I.J. Glycoconj. J. 2008; 25: 121-136Crossref PubMed Scopus (15) Google Scholar, 12Wearne K.A. Winter H.C. O'Shea K. Goldstein I.J. Glycobiology. 2006; 16: 981-990Crossref PubMed Scopus (57) Google Scholar). Lectin microarray is an alternative technology for structural glycomics, where a panel of lectins with various glycan-binding specificities is printed on a microarray, providing a versatile platform for rapid and high throughput analysis of glycan structures without liberation of glycans (13Pilobello K.T. Krishnamoorthy L. Slawek D. Mahal L.K. Chembiochem. 2005; 6: 985-989Crossref PubMed Scopus (244) Google Scholar, 14Kuno A. Uchiyama N. Koseki-Kuno S. Ebe Y. Takashima S. Yamada M. Hirabayashi J. Nat. Methods. 2005; 2: 851-856Crossref PubMed Scopus (433) Google Scholar). Lectins are a class of decoder molecules of cell surface glycans distributed throughout organisms, which mediate various functions through specific glycan recognition. Analytical protocols using lectin microarray have been developed for various sample types: free oligosaccharides (14Kuno A. Uchiyama N. Koseki-Kuno S. Ebe Y. Takashima S. Yamada M. Hirabayashi J. Nat. Methods. 2005; 2: 851-856Crossref PubMed Scopus (433) Google Scholar, 15Uchiyama N. Kuno A. Tateno H. Kubo Y. Mizuno M. Noguchi M. Hirabayashi J. Proteomics. 2008; 8: 3042-3050Crossref PubMed Scopus (51) Google Scholar), tissue sections (16Matsuda A. Kuno A. Ishida H. Kawamoto T. Shoda J. Hirabayashi J. Biochem. Biophys. Res. Commun. 2008; 370: 259-263Crossref PubMed Scopus (58) Google Scholar), cell membrane hydrophobic fractions (17Ebe Y. Kuno A. Uchiyama N. Koseki-Kuno S. Yamada M. Sato T. Narimatsu H. Hirabayashi J. J. Biochem. 2006; 139: 323-327Crossref PubMed Scopus (57) Google Scholar, 18Pilobello K.T. Slawek D.E. Mahal L.K. Proc. Natl. Acad. Sci. U.S.A. 2007; 104: 11534-11539Crossref PubMed Scopus (175) Google Scholar), and even whole cells (19Hsu K.L. Pilobello K.T. Mahal L.K. Nat. Chem. Biol. 2006; 2: 153-157Crossref PubMed Scopus (208) Google Scholar, 20Tateno H. Uchiyama N. Kuno A. Togayachi A. Sato T. Narimatsu H. Hirabayashi J. Glycobiology. 2007; 17: 1138-1146Crossref PubMed Scopus (136) Google Scholar). This technology has just begun to be applied to a wide variety of biological researches, including virus profiling (21Krishnamoorthy L. Bess Jr., J.W. Preston A.B. Nagashima K. Mahal L.K. Nat. Chem. Biol. 2009; 5: 244-250Crossref PubMed Scopus (133) Google Scholar) and cell profiling (17Ebe Y. Kuno A. Uchiyama N. Koseki-Kuno S. Yamada M. Sato T. Narimatsu H. Hirabayashi J. J. Biochem. 2006; 139: 323-327Crossref PubMed Scopus (57) Google Scholar, 20Tateno H. Uchiyama N. Kuno A. Togayachi A. Sato T. Narimatsu H. Hirabayashi J. Glycobiology. 2007; 17: 1138-1146Crossref PubMed Scopus (136) Google Scholar), and development of cancer glycobiomarkers (22Kuno A. Kato Y. Matsuda A. Kaneko M.K. Ito H. Amano K. Chiba Y. Narimatsu H. Hirabayashi J. Mol. Cell Proteomics. 2009; 8: 99-108Abstract Full Text Full Text PDF PubMed Scopus (100) Google Scholar, 23Narimatsu H. Sawaki H. Kuno A. Kaji H. Ito H. Ikehara Y. FEBS J. 2010; 277: 95-105Crossref PubMed Scopus (146) Google Scholar, 24Matsuda A. Kuno A. Kawamoto T. Matsuzaki H. Irimura T. Ikehara Y. Zen Y. Nakanuma Y. M. N. Shoda J. Hirabayashi J. Narimatsu H. 2010; PubMed Scopus Google Scholar). For cell of in hydrophobic fractions are for each analysis H. Kuno A. Y. Hirabayashi J. 2010; PubMed Scopus Google Scholar, A. Y. M. Y. Yamada M. A. Hirabayashi J. J. 2008; Google Scholar). and were to the of the data H. Kuno A. Y. Hirabayashi J. 2010; PubMed Scopus Google Scholar, A. Y. M. Y. Yamada M. A. Hirabayashi J. J. 2008; Google Scholar). we have demonstrated that lectin microarray is also applicable to stem cells M. M. Y. Kuno A. T. Yamada M. H. Y. S. Narimatsu H. Hirabayashi J. A. Cells. 2011; 16: PubMed Scopus Google Scholar, S. Y. Ito Y. Tateno H. M. H. K. Y. Y. H. N. Y. A. Hirabayashi J. K. M. The on Scholar), although we have to a as to how the cellular glycome upon induction of pluripotency. of this technology to the of stem cells has not been we developed an platform of high density lectin microarray with the number of lectins to the glycome for of various stem cell systematic of the cellular glycome was then performed cell types in including iPSCs cell types) and ESCs cell this comprehensive we obtained that all of the four SCs with originally distinct glycan profiles have acquired those similar to ESCs upon induction of pluripotency. also found structural features to iPSCs and which well to the results of gene expression analysis of we the of lectin microarray in the stem cell diagnosis of multiple including between undifferentiated and differentiated cells as well as of the contamination of the