Long-term xeno-free culture of human pluripotent stem cells on hydrogels with optimal elasticity

在具有最佳弹性的水凝胶上长期无异种培养人类多能干细胞

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作者:Akon Higuchi, Shih-Hsuan Kao, Qing-Dong Ling, Yen-Ming Chen, Hsing-Fen Li, Abdullah A Alarfaj, Murugan A Munusamy, Kadarkarai Murugan, Shih-Chang Chang, Hsin-Chung Lee, Shih-Tien Hsu, S Suresh Kumar, Akihiro Umezawa

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.

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