title: A xenogeneic-free system generating functional human gut organoids from pluripotent stem cells.
abstract: Functional intestines are composed of cell types from all 3 primary germ layers and are generated through a highly orchestrated and serial developmental process. Directed differentiation of human pluripotent stem cells (hPSCs) has been shown to yield gut-specific cell types; however, these structures do not reproduce critical functional interactions between cell types of different germ layers. Here, we developed a simple protocol for the generation of mature functional intestinal organoids from hPSCs under xenogeneic-free conditions. The stem cell-derived gut organoids produced here were found to contain distinct types of intestinal cells, including enterocytes, goblet cells, Paneth cells, and enteroendocrine cells, that were derived from all 3 germ layers; moreover, they demonstrated intestinal functions, including peptide absorption, and showed innervated bowel movements in response to stimulation with histamine and anticholinergic drugs. Importantly, the gut organoids obtained using this xenogeneic-free system could be stably maintained in culture for prolonged periods and were successfully engrafted in vivo. Our xenogeneic-free approach for generating gut organoids from hPSCs provides a platform for studying human intestinal diseases and for pharmacological testing.
authors.affiliation: Transplantation Centre, National Centre for Child Health and Development, Tokyo, Japan.|Department of Surgery, Tohoku University, Sendai, Japan.
authors.forename: Hajime
authors.initials: H
authors.lastname: Uchida
authors.affiliation: Department of Reproductive Biology, Centre for Regenerative Medicine, National Research Institute for Child Health and Development, Tokyo, Japan.
authors.forename: Masakazu
authors.initials: M
authors.lastname: Machida
authors.affiliation: Department of Reproductive Biology, Centre for Regenerative Medicine, National Research Institute for Child Health and Development, Tokyo, Japan.
authors.forename: Takumi
authors.initials: T
authors.lastname: Miura
authors.affiliation: Department of Reproductive Biology, Centre for Regenerative Medicine, National Research Institute for Child Health and Development, Tokyo, Japan.
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Kawasaki
authors.affiliation: Life Science Department, Research and Development Center, Dai Nippon Printing Co., Ltd., Tokyo, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Okazaki
authors.affiliation: Transplantation Centre, National Centre for Child Health and Development, Tokyo, Japan.|Department of Surgery, Tohoku University, Sendai, Japan.
authors.forename: Kengo
authors.initials: K
authors.lastname: Sasaki
authors.affiliation: Transplantation Centre, National Centre for Child Health and Development, Tokyo, Japan.
authors.forename: Seisuke
authors.initials: S
authors.lastname: Sakamoto
authors.affiliation: Department of Surgery, Tohoku University, Sendai, Japan.
authors.forename: Noriaki
authors.initials: N
authors.lastname: Ohuchi
authors.affiliation: Transplantation Centre, National Centre for Child Health and Development, Tokyo, Japan.
authors.forename: Mureo
authors.initials: M
authors.lastname: Kasahara
authors.affiliation: Department of Reproductive Biology, Centre for Regenerative Medicine, National Research Institute for Child Health and Development, Tokyo, Japan.
authors.forename: Akihiro
authors.initials: A
authors.lastname: Umezawa
authors.affiliation: Department of Reproductive Biology, Centre for Regenerative Medicine, National Research Institute for Child Health and Development, Tokyo, Japan.|Department of Stem Cell Research, Fukushima Medical University, Fukushima, Japan.
authors.forename: Hidenori
authors.initials: H
authors.lastname: Akutsu
mesh_terms: D002454:Cell Differentiation / Q000235:genetics; D020895:Enterocytes / Q000166:cytology / Q000378:metabolism; D020397:Goblet Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D007422:Intestines / Q000166:cytology*; D050814:Octamer Transcription Factor-3; D009940:Organoids / Q000166:cytology* / Q000378:metabolism; D019879:Paneth Cells / Q000166:cytology / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology*; D055748:SOXB1 Transcription Factors
chemical_list: D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; C490906:SOX1 protein, human; D055748:SOXB1 Transcription Factors