Human PSCs determine the competency of cerebral organoid differentiation via FGF signaling and epigenetic mechanisms

Hirosato Ideno; Kent Imaizumi; Hiroko Shimada; Tsukasa Sanosaka; Akisa Nemoto; Jun Kohyama; Hideyuki Okano

doi:10.1016/j.isci.2022.105140

Human PSCs determine the competency of cerebral organoid differentiation via FGF signaling and epigenetic mechanisms

iScience. 2022 Sep 16;25(10):105140. doi: 10.1016/j.isci.2022.105140. eCollection 2022 Oct 21.

Authors

Hirosato Ideno¹, Kent Imaizumi¹, Hiroko Shimada¹, Tsukasa Sanosaka¹, Akisa Nemoto¹, Jun Kohyama¹, Hideyuki Okano¹

Affiliation

¹ Department of Physiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.

Abstract

Various culture methods have been developed for maintaining human pluripotent stem cells (PSCs). These PSC maintenance methods exhibit biased differentiation; for example, feeder-dependent PSCs efficiently yield cerebral organoids, but it is difficult to generate organoids from feeder-free PSCs. It remains unknown how PSC maintenance conditions affect differentiation. In this study, we identified fibroblast growth factor (FGF) signaling in feeder-free PSC maintenance as a key factor that determines the differentiation toward cerebral organoids. The inhibition of FGF signaling in feeder-free PSCs rescued organoid generation to the same level in feeder-dependent cultures. FGF inhibition induced DNA methylation at the WNT5A locus, and this epigenetic change suppressed the future activation of non-canonical Wnt signaling after differentiation, leading to reliable cerebral organoid generation. This study underscores the importance of PSC culture conditions for directed differentiation into cerebral organoids, and the epigenetic status regulated by FGF signaling is involved in the underlying mechanisms.

Keywords: Cell biology; Genomics; Omics; Stem cells research; Transcriptomics.