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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

1
Department of Orthopedic Surgery, Hokkaido University Graduate School of Medicine, Kita 15, Nishi 7, Kita-ku, Sapporo, Hokkaido 060-8638, Japan
2
Department of Advanced Clinical Glycobiology, Faculty of Medicine and Graduate School of Medicine, Hokkaido, Kita 21, Nishi 11, Kita-ku, Sapporo, Hokkaido 001-0021, Japan
3
Global Station for Soft Matter, Global Institution for Collaborative Research and Education (GSS, GI-CoRE), Hokkaido University, Kita 21, Nishi 11, Kita-ku, Sapporo, Hokkaido 001-0021, Japan
4
Research Center for Glycobiotechnology, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga 525-8577, Japan
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Int. J. Mol. Sci. 2020, 21(1), 231; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21010231
Received: 31 October 2019 / Revised: 26 December 2019 / Accepted: 26 December 2019 / Published: 28 December 2019
(This article belongs to the Special Issue Gangliosides: Modes of Action and Cell Fates)
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 × 104 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. View Full-Text
Keywords: cartilage damage; osteoarthritis; iPSCs; chondrocytes; GSL-glycome analysis; aminolysis-SALSA; tumorigenicity; glycoconjugates cartilage damage; osteoarthritis; iPSCs; chondrocytes; GSL-glycome analysis; aminolysis-SALSA; tumorigenicity; glycoconjugates
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MDPI and ACS Style

Miyazaki, T.; Hanamatsu, H.; Xu, L.; Onodera, T.; Furukawa, J.-i.; Homan, K.; Baba, R.; Kawasaki, T.; Iwasaki, N. Evaluation of Residual Human-Induced Pluripotent Stem Cells in Human Chondrocytes by Cell Type-Specific Glycosphingolipid Glycome Analysis Based on the Aminolysis-SALSA Technique. Int. J. Mol. Sci. 2020, 21, 231. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21010231

AMA Style

Miyazaki T, Hanamatsu H, Xu L, Onodera T, Furukawa J-i, Homan K, Baba R, Kawasaki T, Iwasaki N. Evaluation of Residual Human-Induced Pluripotent Stem Cells in Human Chondrocytes by Cell Type-Specific Glycosphingolipid Glycome Analysis Based on the Aminolysis-SALSA Technique. International Journal of Molecular Sciences. 2020; 21(1):231. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21010231

Chicago/Turabian Style

Miyazaki, Takuji, Hisatoshi Hanamatsu, Liang Xu, Tomohiro Onodera, Jun-ichi Furukawa, Kentaro Homan, Rikiya Baba, Toshisuke Kawasaki, and Norimasa Iwasaki. 2020. "Evaluation of Residual Human-Induced Pluripotent Stem Cells in Human Chondrocytes by Cell Type-Specific Glycosphingolipid Glycome Analysis Based on the Aminolysis-SALSA Technique" International Journal of Molecular Sciences 21, no. 1: 231. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21010231

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