1. Volume 144, Issue 4, Pages 761-770 (April 2013)
Identification of Lineage-Uncommitted, Long-Lived, Label-Retaining Cells in Healthy Human Esophagus and Stomach, and in Metaplastic Esophagus 
Qiuwei Pan, Anna M. Nicholson, Hugh Barr, Lea–Anne Harrison, George D. Wilson, Julia Burkert, Rosemary Jeffery, Malcolm R. Alison, Leendert Looijenga, Wey–Ran Lin, Stuart A.C. McDonald, Nicholas A. Wright, Rebecca Harrison, Maikel P. Peppelenbosch, Janusz A. Jankowski 
Gastroenterology 
Volume 144, Issue 4, Pages (April 2013)
DOI: /j.gastro
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2. Figure 1
Label-retaining cells in the esophageal squamous epithelium. (A) The normal squamous esophageal epithelium. The IBL is highlighted in red, the papillary basal layer (PBL) is highlighted green, with the parabasal layer colored orange and the superficial layer colored brown. (B) Immunohistochemistry showing Ki-67 expression, high-power inset shows expression confined to the basal layer including the IBL and PBL. (C) IdU-positive cells in the squamous epithelium 7 days after infusion, cells are identified in the basal, parabasal, and superficial layers, with high-power image showing an IdU-positive cell in the basal layer. (D) Efflux of cells in the squamous esophagus, counts at 29 days highlight the basal layer as the location of LRCs in the epithelium with most other cells lost from the epithelium.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2013 AGA Institute Terms and Conditions

3. Figure 2
Differential label-retaining cells in the esophageal squamous papillae and interpapillary epithelium. (A) LRCs are identified in the papillary basal layer (PBL) 67 days after infusion with IdU, immunohistochemistry-fluorescence shows 1 positive cell. (B) LRCs (red/555) in the PBL at 67 days co-localizes with cytokeratin (green/488), confirming the epithelial nature of these cells. (C) A LRC (red/555) in the IBL at 67 days resides in the separate yet adjoining location to Ki-67–expressing cells (green/488). (D) There are significantly more LRCs associated with the PBL compared with the IBL (P = .001). (E) LRCs were identified around the papillae within the epithelium. Cells shown here from patient 3 at 11 days postinfusion were seen at the papilla tip.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2013 AGA Institute Terms and Conditions

4. Figure 3
IdU labeling in the human stomach. (A) The gastric unit, where the gland contains chief cells, endocrine cells, and parietal cells. The mid/neck layer is colored orange and the superficial layer is highlighted brown. (B) Immunohistochemistry showing Ki-67 expression, with a high-power inset of expression concentrated in the neck region of the gland. (C) IdU-positive cells identified in the neck and foveolar layers at 7 days after infusion. More IdU-positive cells are seen in the foveolar region, suggesting flux is greater in this direction than to the gland base. (D) IdU-positive cells identified specifically in the neck region of the gland 11 days after infusion, highlighting this as the location of LRCs in the human stomach. (E) LRCs (red/555) in the stomach co-localize with cytokeratin (green/488) at 11 days postinfusion, confirming the epithelial nature of these cells. (F) LRCs in the stomach, which show immunohistochemistry staining of IdU (orange) (red/555) in the neck, reside in a separate yet adjoining location to Ki-67–expressing cells (green/488) at 67 days postinfusion (parietal-type cells with double labeling with IdU-labeled cells).
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2013 AGA Institute Terms and Conditions

5. Figure 4
Label-retaining cells in BE. (A) Schematic of BE. The gland base is highlighted in red, the mid/neck layer is highlighted in orange, and the superficial layer is highlighted in brown. Goblet cells are found within the mid- and superficial layers as shown. (B) Immunohistochemistry showing Ki67 expression, expression is seen toward the base of the glands spreading into the midlayer in these patients. (C) IdU-positive cells in Barrett's 7 days after infusion are identified in the base of the gland (high-power inset shows positive cells) and also in the neck. (D) LRCs are specific to the base of the gland in Barrett's 67 days after infusion of IdU, shown here by immunohistochemistry–horseradish peroxidase using a monoclonal mouse antibody. A positive IdU cell is highlighted. (E) LRCs are specific to the base of the gland in Barrett's 67 days after infusion of IdU, shown here by immunohistochemistry-Fl using a sheep polyclonal antibody. (F) LRCs (red/555) at the base of the Barrett's gland co-localize with cytokeratin (green/488) at 67 days after infusion, confirming their epithelial nature. A positive IdU cell is highlighted.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2013 AGA Institute Terms and Conditions

6. Figure 5
Label-retaining cells in individual cell phenotypes in BE. (A) LRCs (red/555) in the base are located near the Ki-67–positive (green/488) population at 67 days postinfusion. A positive IdU cell is highlighted and shown in the inset. (B) LRCs (red/555) do not express markers consistent with goblet cells (orange/647) or neuroendocrine cells (green/488) in these patients. (C) A LRC (red/555) was identified in the base of the Barrett's Esophagus gland, which co-localizes with chromogranin A (green/488) after 67 days (arrow), in the same gland a LRC adjacent to a neuroendocrine cell can be seen (dashed arrow). (D) LRCs (red/555) in the base of BO glands are not consistent with Paneth cells (green/488) at 67 days, goblet cells stained as a control are seen in this Barrett's segment (orange/647). (E) In this light-field image a segment of Barrett's from patient 4 shows the intestinal stem cell marker LGR5 being expressed in the base of the glands, but not in the associated squamous epithelium. (F) A dark-field image of human LGR5 confirming the signal expression in the base of Barrett's Esophagus glands.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2013 AGA Institute Terms and Conditions

7. Figure 6
Cell counts of IdU-positive cells. (A) A proportion of LRCs are Ki-67 positive at 11 days in the normal squamous epithelium. A total of 20.9% of cells are Ki-67 positive (green bar), with 1.9% of cells confirmed as LRCs (red bar). A small proportion of additional cells at this time have re-entered the cell cycle and are positive for both Ki-67 and IdU (0.21%) (yellow bar). (B) Immunohistochemistry using double staining for IdU (red/555) and Ki-67 (green/488) shows a cell (arrow) in the IBL of the normal squamous epithelium co-labeling for both markers. Ki-67–expressing only cells are seen above. (C) A proportion of LRCs are Ki-67 positive at 11 days in Barrett's glands. A total of 23.43% of cells are Ki-67–positive (green bar), with 2.12% of cells confirmed as LRCs (red bar). A small proportion of cells at this time have re-entered the cell cycle and are positive for both Ki-67 and IdU (0.11%) (yellow bar). (D) Immunohistochemistry using double staining for IdU (red/555) and Ki-67 (green/488) shows 2 neighboring cells within the Barrett's gland co-labeling for both markers. Two further neighboring cells express Ki-67 alone. (E) Counts of IdU-positive cells in the basal layer of the normal squamous epithelium. The number of IdU-positive cells in the basal layer reduces as the chase time increases. At 7 days, 2% of cells retain label whereas by 67 days this is 0.05% of the basal layer. (F) Counts of IdU-positive cells in the base of Barrett's glands. The number of cells labeling in the gland base reduces as the chase time increases up to 67 days. There are 1.4% of cells at the base retaining label at 11 days, but by 67 days only 0.07% are labeled.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2013 AGA Institute Terms and Conditions

8. Supplementary Figure 1
Gastroenterology  , DOI: ( /j.gastro )
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9. Supplementary Figure 2
Gastroenterology  , DOI: ( /j.gastro )
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10. Supplementary Figure 4
Gastroenterology  , DOI: ( /j.gastro )
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