1. Identification and isolation of candidate human colonic clonogenic cells based on cell surface integrin expression 
Koji Fujimoto, R.Daniel Beauchamp, Robert H. Whitehead 
Gastroenterology 
Volume 123, Issue 6, Pages (December 2002)
DOI: /gast
Copyright © 2002 American Gastroenterological Association Terms and Conditions

2. Fig. 1
Expression and distribution of integrin subunits along the crypt and luminal surface in the normal human colon. (A) Frozen sections were stained with primary antibodies to the 12 integrin subunits but only antibodies showing staining are included. Indirect immunofluorescence micrographs from representative fields of the stained sections were taken. Note the strong expression of β1- and α2-integrins at the lower part (one third and two thirds, respectively) of the crypts (original magnification 200×). (B) A second β1-integrin antibody was used to confirm the accuracy of the staining pattern. Both of these antibodies showed the same expression pattern of the β1-integrin subunit in 2 different serial sections (*) (original magnification 200×).
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

3. Fig. 2
Relationship between the expression pattern of β1-integrin by flow cytometry and that in an immunofluorescence study. The cells with higher fluorescence by flow cytometry correspond to the cells located at the lower part of the crypts in the immunofluorescence analysis. The cells in the high fluorescence peak after β1-integrin staining comprised 41% of the cell population.
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

4. Fig. 3
(A) Flow cytometric analysis of isolated single crypt cells stained with integrin antibodies. The dot plot (at upper left side of figure) shows the forward scatter and side scatter of a suspension of isolated crypt cells. This pattern of light scattering was highly reproducible. Note the strong expression of β1- and α2-integrins as compared with the other integrin subunits (horizontal axis: mean fluorescent intensity; longitudinal axis: cell number). Two peaks of fluorescence indicate that the cell population was composed of 2 subpopulations: those with high fluorescence and those with lower fluorescence. The experiments were repeated 3 times by using different human colonic mucosa obtained from the surgical specimens. A representative result is shown. (B) Mean fluorescence intensity of each integrin subunit. □, peak-1 (low intensity); ■, peak-2 (high intensity).
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

5. Fig. 4
Adhesiveness of human crypt cells to ECM proteins. The degree of adhesiveness is represented by the percentage of attached crypt cells. The values are mean ± SD. Three experiments were performed with each ECM protein. The difference of control and ECM proteins was significant for collagen type I, collagen type IV, and laminin (P < 0.05).
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

6. Fig. 5
Colony formation from single cells after cell sorting by using β1-integrin antibody. The colony assay was repeated 5 times by using human crypt cells from different patients. After the isolation of crypt cells, 1 × 105 cells were seeded into each well. Both colonies (>40 cells) and clusters (15–40 cells) were counted after 21 days. Note that the cells that were sorted by the β1-integrin antibody with or without the addition of LIM1863-conditioned medium produced more colonies than the unsorted cells with or without the conditioned medium in all 5 of the assays.
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions