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

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

Research output: Contribution to journalArticlepeer-review

45 Scopus citations

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.

Original languageEnglish
Article number18136
JournalScientific Reports
Volume5
DOIs
StatePublished - 14 Dec 2015

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