Generation of Highly Expandable Intestinal Spheroids Composed of Stem Cells

Ohman Kwon; Won Dong Yu; Ye Seul Son; Kwang Bo Jung; Hana Lee; Mi-Young Son

doi:10.15283/ijsc21209

Generation of Highly Expandable Intestinal Spheroids Composed of Stem Cells

Int J Stem Cells. 2022 Feb 28;15(1):104-111. doi: 10.15283/ijsc21209.

Authors

Ohman Kwon¹, Won Dong Yu^{1

2}, Ye Seul Son¹, Kwang Bo Jung^{1

2}, Hana Lee^{1

2}, Mi-Young Son^{1

2}

Affiliations

¹ Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Korea.
² KRIBB School of Bioscience, Korea University of Science and Technology, Daejeon, Korea.

Abstract

Many of early findings regarding intestinal stem cells (ISCs) and their niche in the human intestine have relied on colorectal cancer cell lines and labor-intensive and time-consuming mouse models. However, these models cannot accurately recapitulate the physiologically relevant aspects of human ISCs. In this study, we demonstrate a reliable and robust culture method for 3D expanding intestinal spheroids (InS^exp) mainly comprising ISCs and progenitors, which can be derived from 3D human intestinal organoids (HIOs). We did functional chararcterization of InS^exp derived from 3D HIOs, differentiated from human pluripotent stem cells, and optimization culture methods. Our results indicate that InS^exp can be rapidly expanded and easily passaged, and show enhanced growth rates via WNT pathway activation. InS^exp are capable of exponential cell expansion and cryopreservation. Furthermore, in vitro-matured HIO-derived InS^exp proliferate faster than immature HIO-derived InS^exp with preservation of the parental HIO characteristics. These findings may facilitate the development of scalable culture systems for the long-term maintenance of human ISCs and provide an alternative platform for studying ISC biology.

Keywords: Expandable Intestinal spheroid (InS^exp); Human intestinal organoid (HIO); Intestinal stem cell; Maturity memory; WNT pathway.