title: A methodological approach to tracing cell lineage in human epithelial tissues.
abstract: Methods for lineage tracing of stem cell progeny in human tissues are currently not available. We describe a technique for detecting the expansion of a single cell's progeny that contain clonal mitochondrial DNA (mtDNA) mutations affecting the expression of mtDNA-encoded cytochrome c oxidase (COX). Because such mutations take up to 40 years to become phenotypically apparent, we believe these clonal patches originate in stem cells. Dual-color enzyme histochemistry was used to identify COX-deficient cells, and mutations were confirmed by microdissection of single cells with polymerase chain reaction sequencing of the entire mtDNA genome. These techniques have been applied to human intestine, liver, pancreas, and skin. Our results suggest that the stem cell niche is located at the base of colonic crypts and above the Paneth cell region in the small intestine, in accord with dynamic cell kinetic studies in animals. In the pancreas, exocrine tissue progenitors appeared to be located in or close to interlobular ducts, and, in the liver, we propose that stem cells are located in the periportal region. In the skin, the origin of a basal cell carcinoma appeared to be from the outer root sheath of the hair follicle. We propose that this is a general method for detecting clonal cell populations from which the location of the niche can be inferred, also affording the generation of cell fate maps, all in human tissues. In addition, the technique allows analysis of the origin of human tumors from specific tissue sites.
authors.affiliation: Centre for Diabetes and Metabolic Medicine, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom.
authors.forename: Tariq G
authors.initials: TG
authors.lastname: Fellous
authors.forename: Stuart A C
authors.initials: SA
authors.lastname: McDonald
authors.forename: Julia
authors.initials: J
authors.lastname: Burkert
authors.forename: Adam
authors.initials: A
authors.lastname: Humphries
authors.forename: Shahriar
authors.initials: S
authors.lastname: Islam
authors.forename: Nemantha M W
authors.initials: NM
authors.lastname: De-Alwis
authors.forename: Lydia
authors.initials: L
authors.lastname: Gutierrez-Gonzalez
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Tadrous
authors.forename: George
authors.initials: G
authors.lastname: Elia
authors.forename: Hemant M
authors.initials: HM
authors.lastname: Kocher
authors.forename: Satyajit
authors.initials: S
authors.lastname: Bhattacharya
authors.forename: Lisa
authors.initials: L
authors.lastname: Mears
authors.forename: Mona
authors.initials: M
authors.lastname: El-Bahrawy
authors.forename: Douglas M
authors.initials: DM
authors.lastname: Turnbull
authors.forename: Robert W
authors.initials: RW
authors.lastname: Taylor
authors.forename: Laura C
authors.initials: LC
authors.lastname: Greaves
authors.forename: Patrick F
authors.initials: PF
authors.lastname: Chinnery
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Day
authors.forename: Nicholas A
authors.initials: NA
authors.lastname: Wright
authors.forename: Malcolm R
authors.initials: MR
authors.lastname: Alison
mesh_terms: D019070:Cell Lineage; D002999:Clone Cells; D004272:DNA, Mitochondrial / Q000235:genetics*; D003576:Electron Transport Complex IV / Q000235:genetics; D004847:Epithelial Cells / Q000166:cytology*; D006801:Humans; D007150:Immunohistochemistry; D009154:Mutation; D055153:Stem Cell Niche / Q000166:cytology
chemical_list: D004272:DNA, Mitochondrial; D003576:Electron Transport Complex IV