title: Long-term xeno-free culture of human pluripotent stem cells on hydrogels with optimal elasticity.
abstract: The tentative clinical application of human pluripotent stem cells (hPSCs), such as human embryonic stem cells and human induced pluripotent stem cells, is restricted by the possibility of xenogenic contamination resulting from the use of mouse embryonic fibroblasts (MEFs) as a feeder layer. Therefore, we investigated hPSC cultures on biomaterials with different elasticities that were grafted with different nanosegments. We prepared dishes coated with polyvinylalcohol-co-itaconic acid hydrogels grafted with an oligopeptide derived from vitronectin (KGGPQVTRGDVFTMP) with elasticities ranging from 10.3 to 30.4 kPa storage moduli by controlling the crosslinking time. The hPSCs cultured on the stiffest substrates (30.4 kPa) tended to differentiate after five days of culture, whereas the hPSCs cultured on the optimal elastic substrates (25 kPa) maintained their pluripotency for over 20 passages under xeno-free conditions. These results indicate that cell culture matrices with optimal elasticity can maintain the pluripotency of hPSCs in culture.
authors.affiliation: Department of Chemical and Materials Engineering, National Central University, No. 300 Jhongli, Taoyuan, 32001 Taiwan.|Department of Reproduction, National Research Institute for Child Health and Development, 2-10-1 Okura, Setagaya-ku, Tokyo 157-8535, Japan.|Nano Medical Engineering Laboratory, RIKEN, 2-1, Hirosawa, Wako, Saitama 351-0198, Japan.|Department of Botany and Microbiology, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia.
authors.forename: Akon
authors.initials: A
authors.lastname: Higuchi
authors.affiliation: Department of Chemical and Materials Engineering, National Central University, No. 300 Jhongli, Taoyuan, 32001 Taiwan.
authors.forename: Shih-Hsuan
authors.initials: SH
authors.lastname: Kao
authors.affiliation: Cathay Medical Research Institute, Cathay General Hospital, No. 32, Ln 160, Jian-Cheng Road, Hsi-Chi City, Taipei, 221, Taiwan.|Graduate Institute of Systems Biology and Bioinformatics, National Central University, No. 300, Jhongda RD., Jhongli, Taoyuan, 32001 Taiwan.
authors.forename: Qing-Dong
authors.initials: QD
authors.lastname: Ling
authors.affiliation: Department of Chemical and Materials Engineering, National Central University, No. 300 Jhongli, Taoyuan, 32001 Taiwan.
authors.forename: Yen-Ming
authors.initials: YM
authors.lastname: Chen
authors.affiliation: Department of Chemical and Materials Engineering, National Central University, No. 300 Jhongli, Taoyuan, 32001 Taiwan.
authors.forename: Hsing-Fen
authors.initials: HF
authors.lastname: Li
authors.affiliation: Department of Botany and Microbiology, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia.
authors.forename: Abdullah A
authors.initials: AA
authors.lastname: Alarfaj
authors.affiliation: Department of Botany and Microbiology, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia.
authors.forename: Murugan A
authors.initials: MA
authors.lastname: Munusamy
authors.affiliation: Division of Entomology, Department of Zoology, School of Life Sciences, Bharathiar University, Coimbatore, Tamil Nadu, India.
authors.forename: Kadarkarai
authors.initials: K
authors.lastname: Murugan
authors.affiliation: Department of Surgery, Cathay General Hospital, No.280, Sec. 4, Ren'ai Rd., Da'an Dist., Taipei, 10693, Taiwan.
authors.forename: Shih-Chang
authors.initials: SC
authors.lastname: Chang
authors.affiliation: Department of Surgery, Cathay General Hospital, No.280, Sec. 4, Ren'ai Rd., Da'an Dist., Taipei, 10693, Taiwan.|Graduate Institute of Translational and Interdisciplinary Medicine, College of Health Science and Technology, National Central University, No. 300, Jhongda RD., Jhongli, Taoyuan, 32001 Taiwan.
authors.forename: Hsin-Chung
authors.initials: HC
authors.lastname: Lee
authors.affiliation: Department of Internal Medicine, Taiwan Landseed Hospital, 77, Kuangtai Road, Pingjen City, Taoyuan 32405, Taiwan.
authors.forename: Shih-Tien
authors.initials: ST
authors.lastname: Hsu
authors.affiliation: Department of Medical Microbiology and Parasitology, Universities Putra Malaysia, Serdang 43400, Slangor, Malaysia.
authors.forename: S Suresh
authors.initials: SS
authors.lastname: Kumar
authors.affiliation: Department of Reproduction, National Research Institute for Child Health and Development, 2-10-1 Okura, Setagaya-ku, Tokyo 157-8535, Japan.
authors.forename: Akihiro
authors.initials: A
authors.lastname: Umezawa
mesh_terms: D000595:Amino Acid Sequence; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D003470:Culture Media / Q000737:chemistry; D055119:Elastic Modulus; D006801:Humans; D020100:Hydrogels / Q000737:chemistry*; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D011142:Polyvinyl Alcohol / Q000737:chemistry; D015203:Reproducibility of Results; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D013386:Succinates / Q000737:chemistry; D013997:Time Factors; D019096:Vitronectin / Q000737:chemistry; D015262:Xenobiotics / Q000737:chemistry
chemical_list: D003470:Culture Media; D020100:Hydrogels; D050814:Octamer Transcription Factor-3; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D013386:Succinates; D019096:Vitronectin; D015262:Xenobiotics; D011142:Polyvinyl Alcohol; C005229:itaconic acid