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Evaluation of Residual Human-Induced Pluripotent Stem Cells in Human Chondrocytes by Cell Type-Specific Glycosphingolipid Glycome Analysis Based on the Aminolysis-SALSA Technique

Cartilage damage may eventually lead to osteoarthritis because it is difficult to repair. Human-induced pluripotent stem cell (iPSC)-derived chondrocytes may potentially be used to treat cartilage damage, but the tumorigenicity of iPSCs is a major concern for their application in regenerative medici...

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Autores principales: Miyazaki, Takuji, Hanamatsu, Hisatoshi, Xu, Liang, Onodera, Tomohiro, Furukawa, Jun-ichi, Homan, Kentaro, Baba, Rikiya, Kawasaki, Toshisuke, Iwasaki, Norimasa
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6982033/
https://www.ncbi.nlm.nih.gov/pubmed/31905707
http://dx.doi.org/10.3390/ijms21010231
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author Miyazaki, Takuji
Hanamatsu, Hisatoshi
Xu, Liang
Onodera, Tomohiro
Furukawa, Jun-ichi
Homan, Kentaro
Baba, Rikiya
Kawasaki, Toshisuke
Iwasaki, Norimasa
author_facet Miyazaki, Takuji
Hanamatsu, Hisatoshi
Xu, Liang
Onodera, Tomohiro
Furukawa, Jun-ichi
Homan, Kentaro
Baba, Rikiya
Kawasaki, Toshisuke
Iwasaki, Norimasa
author_sort Miyazaki, Takuji
collection PubMed
description Cartilage damage may eventually lead to osteoarthritis because it is difficult to repair. Human-induced pluripotent stem cell (iPSC)-derived chondrocytes may potentially be used to treat cartilage damage, but the tumorigenicity of iPSCs is a major concern for their application in regenerative medicine. Many glycoconjugates serve as stem cell markers, and glycosphingolipids (GSLs) including H type 1 antigen (Fucα1-2Galβ1-3GlcNAc) have been expressed on the surface of iPSCs. The purpose of the present study was to investigate whether GSL-glycome analysis is useful for quality control of residual iPSCs in chondrocytes. We performed GSL-glycome analysis of undifferentiated iPSCs in chondrocytes by combining glycoblotting and aminolysis-sialic acid linkage-specific alkylamidation (SALSA) method, enabling the detection of small quantities of iPSC-specific GSL-glycans from 5 × 10(4) cells. Furthermore, we estimated the residual amount of iPSCs using R-17F antibody, which possesses cytotoxic activity toward iPSCs that is dependent on the Lacto-N-fucopentaose I (LNFP I) of GSL. Moreover, we could detect a small number of LNFP I during mesenchymal stem cells (MSCs) differentiation from iPSCs. This is the first demonstration that GSL-glycome analysis is useful for detecting undifferentiated iPSCs, and can thereby support safe regenerative medicine.
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spelling pubmed-69820332020-02-07 Evaluation of Residual Human-Induced Pluripotent Stem Cells in Human Chondrocytes by Cell Type-Specific Glycosphingolipid Glycome Analysis Based on the Aminolysis-SALSA Technique Miyazaki, Takuji Hanamatsu, Hisatoshi Xu, Liang Onodera, Tomohiro Furukawa, Jun-ichi Homan, Kentaro Baba, Rikiya Kawasaki, Toshisuke Iwasaki, Norimasa Int J Mol Sci Article Cartilage damage may eventually lead to osteoarthritis because it is difficult to repair. Human-induced pluripotent stem cell (iPSC)-derived chondrocytes may potentially be used to treat cartilage damage, but the tumorigenicity of iPSCs is a major concern for their application in regenerative medicine. Many glycoconjugates serve as stem cell markers, and glycosphingolipids (GSLs) including H type 1 antigen (Fucα1-2Galβ1-3GlcNAc) have been expressed on the surface of iPSCs. The purpose of the present study was to investigate whether GSL-glycome analysis is useful for quality control of residual iPSCs in chondrocytes. We performed GSL-glycome analysis of undifferentiated iPSCs in chondrocytes by combining glycoblotting and aminolysis-sialic acid linkage-specific alkylamidation (SALSA) method, enabling the detection of small quantities of iPSC-specific GSL-glycans from 5 × 10(4) cells. Furthermore, we estimated the residual amount of iPSCs using R-17F antibody, which possesses cytotoxic activity toward iPSCs that is dependent on the Lacto-N-fucopentaose I (LNFP I) of GSL. Moreover, we could detect a small number of LNFP I during mesenchymal stem cells (MSCs) differentiation from iPSCs. This is the first demonstration that GSL-glycome analysis is useful for detecting undifferentiated iPSCs, and can thereby support safe regenerative medicine. MDPI 2019-12-28 /pmc/articles/PMC6982033/ /pubmed/31905707 http://dx.doi.org/10.3390/ijms21010231 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Miyazaki, Takuji
Hanamatsu, Hisatoshi
Xu, Liang
Onodera, Tomohiro
Furukawa, Jun-ichi
Homan, Kentaro
Baba, Rikiya
Kawasaki, Toshisuke
Iwasaki, Norimasa
Evaluation of Residual Human-Induced Pluripotent Stem Cells in Human Chondrocytes by Cell Type-Specific Glycosphingolipid Glycome Analysis Based on the Aminolysis-SALSA Technique
title Evaluation of Residual Human-Induced Pluripotent Stem Cells in Human Chondrocytes by Cell Type-Specific Glycosphingolipid Glycome Analysis Based on the Aminolysis-SALSA Technique
title_full Evaluation of Residual Human-Induced Pluripotent Stem Cells in Human Chondrocytes by Cell Type-Specific Glycosphingolipid Glycome Analysis Based on the Aminolysis-SALSA Technique
title_fullStr Evaluation of Residual Human-Induced Pluripotent Stem Cells in Human Chondrocytes by Cell Type-Specific Glycosphingolipid Glycome Analysis Based on the Aminolysis-SALSA Technique
title_full_unstemmed Evaluation of Residual Human-Induced Pluripotent Stem Cells in Human Chondrocytes by Cell Type-Specific Glycosphingolipid Glycome Analysis Based on the Aminolysis-SALSA Technique
title_short Evaluation of Residual Human-Induced Pluripotent Stem Cells in Human Chondrocytes by Cell Type-Specific Glycosphingolipid Glycome Analysis Based on the Aminolysis-SALSA Technique
title_sort evaluation of residual human-induced pluripotent stem cells in human chondrocytes by cell type-specific glycosphingolipid glycome analysis based on the aminolysis-salsa technique
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6982033/
https://www.ncbi.nlm.nih.gov/pubmed/31905707
http://dx.doi.org/10.3390/ijms21010231
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