Transcriptomic analysis of feeder-free culture system for maintaining naïve-state pluripotency in human pluripotent stem cells

Wataru Isono; Tomoyuki Kawasaki; Justin K Ichida; Kazunori Nagasaka; Osamu Hiraike; Akihiro Umezawa; Hidenori Akutsu

doi:10.21037/sci-2022-043

Transcriptomic analysis of feeder-free culture system for maintaining naïve-state pluripotency in human pluripotent stem cells

Stem Cell Investig. 2023 Apr 19:10:10. doi: 10.21037/sci-2022-043. eCollection 2023.

Authors

Wataru Isono^{1

2}, Tomoyuki Kawasaki¹, Justin K Ichida³, Kazunori Nagasaka², Osamu Hiraike⁴, Akihiro Umezawa¹, Hidenori Akutsu¹

Affiliations

¹ Center for Regenerative Medicine, National Center for Child Health and Development, Setagaya, Tokyo, Japan.
² Department of Obstetrics and Gynecology, Teikyo University School of Medicine, Itabashi, Tokyo, Japan.
³ Department of Stem Cell Biology and Regenerative Medicine, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA.
⁴ Department of Obstetrics and Gynaecology, Graduate School of Medicine, The University of Tokyo, Bunkyo, Tokyo, Japan.

Abstract

Background: Human pluripotent stem cells (hPSCs) such as embryonic stem cells (ESCs) and induced pluripotent stem cells (PSCs) have the capacity of self-renewal and multilineage differentiation in vitro. Conventional hPSCs, which are in a primed state, can produce various types of differentiated cells. However, the variability in their degree of pluripotency and differentiation propensities, which is influenced by the inductive methods and culture conditions, limit their availability. Therefore, PSCs in a naïve state are a promising source of PSCs.

Methods: We recently developed a culture system for naïve hPSCs using an inhibitor of the NOTCH signaling pathway and a histone H3 methyltransferase disruptor. This culture system requires feeder cells for stably maintaining the naïve hPSCs. We aimed to develop a culture system for hPSCs that could maintain pluripotency under feeder-free conditions.

Results: We used two inhibitors to develop an alternative feeder-free culture system to obtain naïve hPSCs. The naïve cells underwent stable cellular proliferation and were positive for naïve stem cell markers; in addition, they could differentiate into the three germ layers. These feeder-free dome-shaped induced pluripotent stem cells (FFDS-iPSCs) have characteristics similar to that of naïve-like PSCs.

Conclusions: The naive hPSCs under feeder-free conditions could ensure supply of cells for various applications in regenerative medicine and disease modeling.

Keywords: Induced pluripotent stem cells (iPS cells); naïve stem cells; regenerative medicine.