Xeno-free culture of human pluripotent stem cells on oligopeptide-grafted hydrogels with various molecular designs

人类多能干细胞在具有不同分子设计的寡肽接枝水凝胶上的无异种培养

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作者:Yen-Ming Chen, Li-Hua Chen, Meng-Pei Li, Hsing-Fen Li, Akon Higuchi, S Suresh Kumar, Qing-Dong Ling, Abdullah A Alarfaj, Murugan A Munusamy, Yung Chang, Giovanni Benelli, Kadarkarai Murugan, Akihiro Umezawa

Abstract

Establishing cultures of human embryonic (ES) and induced pluripotent (iPS) stem cells in xeno-free conditions is essential for producing clinical-grade cells. Development of cell culture biomaterials for human ES and iPS cells is critical for this purpose. We designed several structures of oligopeptide-grafted poly (vinyl alcohol-co-itaconic acid) hydrogels with optimal elasticity, and prepared them in formations of single chain, single chain with joint segment, dual chain with joint segment, and branched-type chain. Oligopeptide sequences were selected from integrin- and glycosaminoglycan-binding domains of the extracellular matrix. The hydrogels grafted with vitronectin-derived oligopeptides having a joint segment or a dual chain, which has a storage modulus of 25 kPa, supported the long-term culture of human ES and iPS cells for over 10 passages. The dual chain and/or joint segment with cell adhesion molecules on the hydrogels facilitated the proliferation and pluripotency of human ES and iPS cells.

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