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.
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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

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Figure 4

SP cells have stem cell properties in clonogenic and in 3D organotypic culture assays.

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SP cells have stem cell properties in clonogenic and in 3D organotypic c...
(A) When SP, NSP, and unsorted control cells were grown for 4 weeks, there were 34.33 ± 2.84 colonies with 1,000 SP cells and 64 ± 4.93 colonies with 2,500 SP cells (n = 3 experiments), compared with absent colonies with control or NSP cells (n = 3 experiments). Arrowheads indicate colonies. *P < 0.001 compared with NSP. (B) Two-photon microscopy of the disordered cells on the organotypic culture (day 12) after seeding 103 NSP cells isolated from GFP+ mice (green) with DAPI+ nuclei (blue); top view, z-stack, 53.8 μm in 0.78-μm increments. (C) Two-photon microscopy of the fully formed epithelium on day 12 after seeding 103 SP cells isolated from GFP+ mice (green) with DAPI+ nuclei (blue); top view, z-stack, 60 μm in 1-μm increments. Arrowheads indicate basement membrane. (D and E) Unsorted GFP– cells (3 × 104) did not form a complete epithelium. (F) GFP– immunohistochemistry and (G) DAPI+ nuclei. (H and I) Sorted 103 NSP cells from GFP+ mice mixed with 3 × 104 unsorted GFP– cells did not form a complete epithelium. (J) Mosaic GFP+ pattern and (K) DAPI+ nuclei. (L and M) SP cells (103) from GFP+ mice mixed with 3 × 104 unsorted GFP– cells. There was complete epithelial formation with keratinization. (N) GFP+ immunohistochemistry and (O) DAPI+ nuclei. Dotted line in B–O indicates basement membrane. Scale bars: 5 mm (A), 25 μm (B–O). Original magnification, ×20 (A); ×100 (D, H, and L); ×200 (B and C); ×400 (E–G, I–K, and M–O).