title: Functional engraftment of colon epithelium expanded in vitro from a single adult Lgr5⁺ stem cell.
abstract: Adult stem-cell therapy holds promise for the treatment of gastrointestinal diseases. Here we describe methods for long-term expansion of colonic stem cells positive for leucine-rich repeat containing G protein-coupled receptor 5 (Lgr5(+) cells) in culture. To test the transplantability of these cells, we reintroduced cultured GFP(+) colon organoids into superficially damaged mouse colon. The transplanted donor cells readily integrated into the mouse colon, covering the area that lacked epithelium as a result of the introduced damage in recipient mice. At 4 weeks after transplantation, the donor-derived cells constituted a single-layered epithelium, which formed self-renewing crypts that were functionally and histologically normal. Moreover, we observed long-term (>6 months) engraftment with transplantation of organoids derived from a single Lgr5(+) colon stem cell after extensive in vitro expansion. These data show the feasibility of colon stem-cell therapy based on the in vitro expansion of a single adult colonic stem cell.
authors.affiliation: Department of Gastroenterology and Hepatology, Graduate School, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo, Japan.
authors.forename: Shiro
authors.initials: S
authors.lastname: Yui
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Nakamura
authors.forename: Toshiro
authors.initials: T
authors.lastname: Sato
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Nemoto
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Mizutani
authors.forename: Xiu
authors.initials: X
authors.lastname: Zheng
authors.forename: Shizuko
authors.initials: S
authors.lastname: Ichinose
authors.forename: Takashi
authors.initials: T
authors.lastname: Nagaishi
authors.forename: Ryuichi
authors.initials: R
authors.lastname: Okamoto
authors.forename: Kiichiro
authors.initials: K
authors.lastname: Tsuchiya
authors.forename: Hans
authors.initials: H
authors.lastname: Clevers
authors.forename: Mamoru
authors.initials: M
authors.lastname: Watanabe
mesh_terms: D053687:Adult Stem Cells / Q000378:metabolism*; D000818:Animals; D001835:Body Weight / Q000502:physiology; D002478:Cells, Cultured; D053379:Chromogranin A / Q000378:metabolism; D003106:Colon / Q000166:cytology* / Q000637:transplantation*; D051545:Cyclooxygenase 1 / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000172:deficiency; D004848:Epithelium / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000235:genetics; D066298:In Vitro Techniques; D019394:Ki-67 Antigen / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D009940:Organoids / Q000166:cytology / Q000502:physiology; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism*; D012235:Rhodamines / Q000378:metabolism; D013997:Time Factors; D019737:Transplants
chemical_list: D053379:Chromogranin A; D004268:DNA-Binding Proteins; D019394:Ki-67 Antigen; C493597:Lgr5 protein, mouse; D008565:Membrane Proteins; C499308:Rag2 protein, mouse; D043562:Receptors, G-Protein-Coupled; D012235:Rhodamines; C506155:chromogranin A, mouse; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C009434:tetramethylrhodamine isothiocyanate; D051545:Cyclooxygenase 1; C496512:Ptgs1 protein, mouse