title: Cloning and variation of ground state intestinal stem cells.
abstract: Stem cells of the gastrointestinal tract, pancreas, liver and other columnar epithelia collectively resist cloning in their elemental states. Here we demonstrate the cloning and propagation of highly clonogenic, 'ground state' stem cells of the human intestine and colon. We show that derived stem-cell pedigrees sustain limited copy number and sequence variation despite extensive serial passaging and display exquisitely precise, cell-autonomous commitment to epithelial differentiation consistent with their origins along the intestinal tract. This developmentally patterned and epigenetically maintained commitment of stem cells is likely to enforce the functional specificity of the adult intestinal tract. Using clonally derived colonic epithelia, we show that toxins A or B of the enteric pathogen Clostridium difficile recapitulate the salient features of pseudomembranous colitis. The stability of the epigenetic commitment programs of these stem cells, coupled with their unlimited replicative expansion and maintained clonogenicity, suggests certain advantages for their use in disease modelling and regenerative medicine.
authors.affiliation: The Jackson Laboratory for Genomic Medicine, Farmington, Connecticut 06032, USA.
authors.forename: Xia
authors.initials: X
authors.lastname: Wang
authors.affiliation: The Jackson Laboratory for Genomic Medicine, Farmington, Connecticut 06032, USA.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Yamamoto
authors.affiliation: 1] The Jackson Laboratory for Genomic Medicine, Farmington, Connecticut 06032, USA [2] Department of Genetics and Developmental Biology, University of Connecticut Health Center, Farmington, Connecticut 06032, USA.
authors.forename: Lane H
authors.initials: LH
authors.lastname: Wilson
authors.affiliation: Genome Institute of Singapore, Agency for Science, Technology and Research, 138672 Singapore.
authors.forename: Ting
authors.initials: T
authors.lastname: Zhang
authors.affiliation: Department of Pathology, Brigham and Women's Hospital, Boston, Massachusetts 02118, USA.
authors.forename: Brooke E
authors.initials: BE
authors.lastname: Howitt
authors.affiliation: Department of Pathology, Microbiology, and Immunology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA.
authors.forename: Melissa A
authors.initials: MA
authors.lastname: Farrow
authors.affiliation: Genome Institute of Singapore, Agency for Science, Technology and Research, 138672 Singapore.
authors.forename: Florian
authors.initials: F
authors.lastname: Kern
authors.affiliation: The Jackson Laboratory for Genomic Medicine, Farmington, Connecticut 06032, USA.
authors.forename: Gang
authors.initials: G
authors.lastname: Ning
authors.affiliation: The Jackson Laboratory for Genomic Medicine, Farmington, Connecticut 06032, USA.
authors.forename: Yue
authors.initials: Y
authors.lastname: Hong
authors.affiliation: 1] Genome Institute of Singapore, Agency for Science, Technology and Research, 138672 Singapore [2] Department of Ophthalmology, Yong Loo Lin School of Medicine, National University of Singapore, 119228 Singapore.
authors.forename: Chiea Chuen
authors.initials: CC
authors.lastname: Khor
authors.affiliation: The Jackson Laboratory for Genomic Medicine, Farmington, Connecticut 06032, USA.
authors.forename: Benoit
authors.initials: B
authors.lastname: Chevalier
authors.affiliation: Genome Institute of Singapore, Agency for Science, Technology and Research, 138672 Singapore.
authors.forename: Denis
authors.initials: D
authors.lastname: Bertrand
authors.affiliation: Genome Institute of Singapore, Agency for Science, Technology and Research, 138672 Singapore.
authors.forename: Lingyan
authors.initials: L
authors.lastname: Wu
authors.affiliation: Genome Institute of Singapore, Agency for Science, Technology and Research, 138672 Singapore.
authors.forename: Niranjan
authors.initials: N
authors.lastname: Nagarajan
authors.affiliation: Department of Pediatrics, Division of Gastroenterology, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.
authors.forename: Francisco A
authors.initials: FA
authors.lastname: Sylvester
authors.affiliation: Division of Digestive Diseases, Hepatology, and Nutrition, Connecticut Children's Medical Center, Hartford, Connecticut 06106, USA.
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Hyams
authors.affiliation: Department of Medicine, University of Connecticut Health Center, Farmington, Connecticut 06032, USA.
authors.forename: Thomas
authors.initials: T
authors.lastname: Devers
authors.affiliation: Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Roderick
authors.initials: R
authors.lastname: Bronson
authors.affiliation: Department of Pathology, Microbiology, and Immunology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA.
authors.forename: D Borden
authors.initials: DB
authors.lastname: Lacy
authors.affiliation: Department of Medicine, National University of Singapore, 119228 Singapore.
authors.forename: Khek Yu
authors.initials: KY
authors.lastname: Ho
authors.affiliation: Department of Pathology, Brigham and Women's Hospital, Boston, Massachusetts 02118, USA.
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Crum
authors.affiliation: 1] The Jackson Laboratory for Genomic Medicine, Farmington, Connecticut 06032, USA [2] Genome Institute of Singapore, Agency for Science, Technology and Research, 138672 Singapore [3] Department of Medicine, National University of Singapore, 119228 Singapore [4] Multiclonal Therapeutics, Inc., Farmington, Connecticut 06032, USA.
authors.forename: Frank
authors.initials: F
authors.lastname: McKeon
authors.affiliation: 1] The Jackson Laboratory for Genomic Medicine, Farmington, Connecticut 06032, USA [2] Department of Genetics and Developmental Biology, University of Connecticut Health Center, Farmington, Connecticut 06032, USA [3] Department of Pathology, Brigham and Women's Hospital, Boston, Massachusetts 02118, USA [4] Department of Medicine, National University of Singapore, 119228 Singapore [5] Multiclonal Therapeutics, Inc., Farmington, Connecticut 06032, USA.
authors.forename: Wa
authors.initials: W
authors.lastname: Xian
mesh_terms: D001427:Bacterial Toxins / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage; D002478:Cells, Cultured; D002999:Clone Cells / Q000166:cytology / Q000378:metabolism; D016360:Clostridioides difficile / Q000502:physiology; D003106:Colon / Q000166:cytology / Q000187:drug effects; D004761:Enterocolitis, Pseudomembranous / Q000382:microbiology / Q000473:pathology; D044127:Epigenesis, Genetic / Q000235:genetics; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism; D005333:Fetus / Q000166:cytology; D042822:Genomic Instability / Q000235:genetics; D006801:Humans; D007421:Intestine, Small / Q000166:cytology; D007422:Intestines / Q000166:cytology* / Q000187:drug effects; D009940:Organoids / Q000166:cytology / Q000254:growth & development; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*
chemical_list: D001427:Bacterial Toxins; C488583:Clostridium difficile lethal toxin B