A subpopulation of mouse esophageal basal cells has properties of stem cells with the capacity for self-renewal and lineage specification
Jiri Kalabis, … , Meenhard Herlyn, Anil K. Rustgi
Jiri Kalabis, … , Meenhard Herlyn, Anil K. Rustgi
Published November 6, 2008
Citation Information: J Clin Invest. 2008;118(12):3860-3869. https://doi.org/10.1172/JCI35012.
View: Text | PDF
Research Article

A subpopulation of mouse esophageal basal cells has properties of stem cells with the capacity for self-renewal and lineage specification

Abstract

The esophageal epithelium is a prototypical stratified squamous epithelium that exhibits an exquisite equilibrium between proliferation and differentiation. After basal cells proliferate, they migrate outward toward the luminal surface, undergo differentiation, and eventually slough due to apoptosis. The identification and characterization of stem cells responsible for the maintenance of the esophageal epithelium remains elusive. Here, we employed Hoechst dye extrusion and BrdU label–retaining assays to identify in mice a potential esophageal stem cell population that localizes to the basal cell compartment. The self-renewing capacity of this population was characterized using a clonogenic assay and a 3D organotypic culture model. The putative esophageal stem cells were also capable of epithelial reconstitution in vivo in direct esophageal epithelial injury models. In both the 3D organotypic culture and direct mucosal injury models, the putative stem cells gave rise to undifferentiated and differentiated cells. These studies therefore provide a basis for understanding the regenerative capacity and biology of the esophageal epithelium when it is faced with injurious insults.

Authors

Jiri Kalabis, Kenji Oyama, Takaomi Okawa, Hiroshi Nakagawa, Carmen Z. Michaylira, Douglas B. Stairs, Jose-Luiz Figueiredo, Umar Mahmood, J. Alan Diehl, Meenhard Herlyn, Anil K. Rustgi

×

Figure 7

CD34+GFP+ cells have the capacity to contribute to esophageal epithelial restitution after induction of injury.

Options: View larger image (or click on image) Download as PowerPoint
CD34+GFP+ cells have the capacity to contribute to esophageal epithelial...
(A) CD34+ (green) or CD34– (red) esophageal epithelial cells were CK+ (94.8% ± 1.66% or 91.24% ± 1.38% by FACS, respectively; data not shown), and nuclei were stained with DAPI (blue). Original magnification, ×1,000. Scale bars: 5 μm. (B) Comparison of selected markers (CD34, ABCG2, EphA3) in SP cells normalized to NSP cells by RT-PCR (*P < 0.01 compared with NSP; n = 3 experiments). (C) Comparison of selected markers in CD34+ cells normalized to CD34– cells by RT-PCR (*P < 0.01 compared with CD34–; n = 3 experiments). In B and C, data are mean ± SEM. (D, G, and J) PBS (control) was injected in the submucosa after mucosal injury. The epithelium reformed after 48 hours. There were no GFP+ cells (red) in PBS-injected tissues. (E, H, and K) CD34–GFP+ cells (3 × 104) were injected into the submucosa after induction of mucosal injury. The epithelium re-formed after 48 hours. There were few GFP+ cells (red) in the epithelium. (F, I, and L) CD34+GFP+ cells (3 × 104) were injected into the submucosa after induction of mucosal injury. The epithelium re-formed after 48 hours. The re-formed epithelium was GFP+, consistent with the migration of stem cells and emergence of differentiated lineages. Some GFP+ cells remained in the submucosa. Arrowheads indicate basement membrane. Arrows indicate site of cell injection in the submucosa. Original magnification, ×40 (D–F); ×100 (G–L). Scale bars: 25 μm.