Distinct human stem cell populations in small and large intestine.
The intestine is composed of an epithelial layer containing rapidly proliferating cells that mature into two regions, the small and the large intestine. Although previous studies have identified stem cells as the cell-of-origin for intestinal epithelial cells, no studies have directly compared stem cells derived from these anatomically distinct regions. Here, we examine intrinsic differences between primary epithelial cells isolated from human fetal small and large intestine, after in vitro expansion, using the Wnt agonist R-spondin 2. We utilized flow cytometry, fluorescence-activated cell sorting, gene expression analysis and a three-dimensional in vitro differentiation assay to characterize their stem cell properties. We identified stem cell markers that separate subpopulations of colony-forming cells in the small and large intestine and revealed important differences in differentiation, proliferation and disease pathways using gene expression analysis. Single cells from small and large intestine cultures formed organoids that reflect the distinct cellular hierarchy found in vivo and respond differently to identical exogenous cues. Our characterization identified numerous differences between small and large intestine epithelial stem cells suggesting possible connections to intestinal disease.
Department of Pathology, University of Pittsburgh School of Medicine, 200 Lothrop Street, Pittsburgh, PA, 15261, United States of America; McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA, 15219, United States of America.
Julie M
JM
Cramer
Department of Bioengineering, University of Pittsburgh, 3700 O'Hara St, Pittsburgh, PA, 15261, United States of America.
Timothy
T
Thompson
Department of Pathology, University of Pittsburgh, Shadyside Hospital, West Wing, WG 02.11, 5230 Center Avenue, Pittsburgh, PA 15232, United States of America.
Albert
A
Geskin
Department of Pathology, University of Pittsburgh, Shadyside Hospital, West Wing, WG 02.11, 5230 Center Avenue, Pittsburgh, PA 15232, United States of America.
William
W
LaFramboise
Department of Pathology, University of Pittsburgh School of Medicine, 200 Lothrop Street, Pittsburgh, PA, 15261, United States of America; McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA, 15219, United States of America.
Eric
E
Lagasse
D015415:Biomarkers / Q000378:metabolism*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation; D002478:Cells, Cultured; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology*; D007420:Intestine, Large / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D007421:Intestine, Small / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000502:physiology
D015415:Biomarkers; D036341:Intercellular Signaling Peptides and Proteins