Open AccessA Methodological Approach to Tracing Cell Lineage in Human Epithelial Tissues
Author(s) -
Fellous Tariq G.,
McDonald Stuart A.C.,
Burkert Julia,
Humphries Adam,
Islam Shahriar,
DeAlwis Nemantha M.W.,
GutierrezGonzalez Lydia,
Tadrous Paul J.,
Elia George,
Kocher Hemant M.,
Bhattacharya Satyajit,
Mears Lisa,
ElBahrawy Mona,
Turnbull Douglas M.,
Taylor Robert W.,
Greaves Laura C.,
Chinnery Patrick F.,
Day Christopher P.,
Wright Nicholas A.,
Alison Malcolm R.
Publication year - 2009
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1002/stem.67
Subject(s) - biology , stem cell , microdissection , mitochondrial dna , progenitor cell , pancreas , cell , lineage (genetic) , microbiology and biotechnology , genetics , gene , biochemistry
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. S TEM C ELLS 2009;27:1410–1420