1. Volume 115, Issue 3, Pages 605-617 (September 1998)
Identification of distinct molecular phenotypes in cultured gastrointestinal smooth muscle cells 
Jacqueline Brittingham, Christopher Phiel, Wendy C. Trzyna, Vijayalakshmi Gabbeta, Kirk M. McHugh 
Gastroenterology 
Volume 115, Issue 3, Pages (September 1998)
DOI: /S (98)
Copyright © 1998 American Gastroenterological Association Terms and Conditions

2. Fig. 1
Phase-contrast light micrographs of cultured ISMCs plated at low density. (A) Day 1–cultured ISMCs show a broad, spread shape with numerous mitotic figures present. (B) Day 3–cultured ISMCs show a similar shape as day 1 cells with an increase in cell-to-cell contact. (C) Day 5–cultured ISMCs show a more fusiform cellular shape with regions of cells aligned in parallel. (D) Day 7–cultured ISMCs appear subconfluent and show a fusiform shape with regions of cells aligned in close, parallel orientation. (E) Day 10–cultured ISMCs show a regular fusiform shape with overlapping cell boundaries forming broad sheets of cells aligned in close, parallel orientation. (F) Day 21–cultured ISMCs show a hills-and-valley shape (H and V) with numerous areas of cellular retraction within the valleys.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

3. Fig. 1
Phase-contrast light micrographs of cultured ISMCs plated at low density. (A) Day 1–cultured ISMCs show a broad, spread shape with numerous mitotic figures present. (B) Day 3–cultured ISMCs show a similar shape as day 1 cells with an increase in cell-to-cell contact. (C) Day 5–cultured ISMCs show a more fusiform cellular shape with regions of cells aligned in parallel. (D) Day 7–cultured ISMCs appear subconfluent and show a fusiform shape with regions of cells aligned in close, parallel orientation. (E) Day 10–cultured ISMCs show a regular fusiform shape with overlapping cell boundaries forming broad sheets of cells aligned in close, parallel orientation. (F) Day 21–cultured ISMCs show a hills-and-valley shape (H and V) with numerous areas of cellular retraction within the valleys.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

4. Fig. 1
Phase-contrast light micrographs of cultured ISMCs plated at low density. (A) Day 1–cultured ISMCs show a broad, spread shape with numerous mitotic figures present. (B) Day 3–cultured ISMCs show a similar shape as day 1 cells with an increase in cell-to-cell contact. (C) Day 5–cultured ISMCs show a more fusiform cellular shape with regions of cells aligned in parallel. (D) Day 7–cultured ISMCs appear subconfluent and show a fusiform shape with regions of cells aligned in close, parallel orientation. (E) Day 10–cultured ISMCs show a regular fusiform shape with overlapping cell boundaries forming broad sheets of cells aligned in close, parallel orientation. (F) Day 21–cultured ISMCs show a hills-and-valley shape (H and V) with numerous areas of cellular retraction within the valleys.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

5. Fig. 1
Phase-contrast light micrographs of cultured ISMCs plated at low density. (A) Day 1–cultured ISMCs show a broad, spread shape with numerous mitotic figures present. (B) Day 3–cultured ISMCs show a similar shape as day 1 cells with an increase in cell-to-cell contact. (C) Day 5–cultured ISMCs show a more fusiform cellular shape with regions of cells aligned in parallel. (D) Day 7–cultured ISMCs appear subconfluent and show a fusiform shape with regions of cells aligned in close, parallel orientation. (E) Day 10–cultured ISMCs show a regular fusiform shape with overlapping cell boundaries forming broad sheets of cells aligned in close, parallel orientation. (F) Day 21–cultured ISMCs show a hills-and-valley shape (H and V) with numerous areas of cellular retraction within the valleys.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

6. Fig. 1
Phase-contrast light micrographs of cultured ISMCs plated at low density. (A) Day 1–cultured ISMCs show a broad, spread shape with numerous mitotic figures present. (B) Day 3–cultured ISMCs show a similar shape as day 1 cells with an increase in cell-to-cell contact. (C) Day 5–cultured ISMCs show a more fusiform cellular shape with regions of cells aligned in parallel. (D) Day 7–cultured ISMCs appear subconfluent and show a fusiform shape with regions of cells aligned in close, parallel orientation. (E) Day 10–cultured ISMCs show a regular fusiform shape with overlapping cell boundaries forming broad sheets of cells aligned in close, parallel orientation. (F) Day 21–cultured ISMCs show a hills-and-valley shape (H and V) with numerous areas of cellular retraction within the valleys.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

7. Fig. 1
Phase-contrast light micrographs of cultured ISMCs plated at low density. (A) Day 1–cultured ISMCs show a broad, spread shape with numerous mitotic figures present. (B) Day 3–cultured ISMCs show a similar shape as day 1 cells with an increase in cell-to-cell contact. (C) Day 5–cultured ISMCs show a more fusiform cellular shape with regions of cells aligned in parallel. (D) Day 7–cultured ISMCs appear subconfluent and show a fusiform shape with regions of cells aligned in close, parallel orientation. (E) Day 10–cultured ISMCs show a regular fusiform shape with overlapping cell boundaries forming broad sheets of cells aligned in close, parallel orientation. (F) Day 21–cultured ISMCs show a hills-and-valley shape (H and V) with numerous areas of cellular retraction within the valleys.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

8. Fig. 2
(A) Northern blot analysis of cultured ISMCs plated on plastic at low density/10% FBS probed with the γ-smooth muscle, α-smooth muscle, and β-cytoplasmic and γ-cytoplasmic isoactins at isolated muscle strips (MS), immediately after enzyme dissociation (0), and at days 1, 3, 5, 7, and 10 of culture. (B) Relative expression of γ-smooth muscle (●), α-smooth muscle (○), β-cytoplasmic (□), and γ-cytoplasmic (■) isoactins in isolated muscle strips, immediately after enzyme dissociation (0), and at days 1, 3, 5, 7, and 10 of culture. Quantitation values have been normalized to GAPDH expression (not shown).
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

9. Fig. 2
(A) Northern blot analysis of cultured ISMCs plated on plastic at low density/10% FBS probed with the γ-smooth muscle, α-smooth muscle, and β-cytoplasmic and γ-cytoplasmic isoactins at isolated muscle strips (MS), immediately after enzyme dissociation (0), and at days 1, 3, 5, 7, and 10 of culture. (B) Relative expression of γ-smooth muscle (●), α-smooth muscle (○), β-cytoplasmic (□), and γ-cytoplasmic (■) isoactins in isolated muscle strips, immediately after enzyme dissociation (0), and at days 1, 3, 5, 7, and 10 of culture. Quantitation values have been normalized to GAPDH expression (not shown).
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

10. Fig. 3
(A) Northern blot analysis of cultured ISMCs plated at high density (HD)/10% FBS, high density/0.5% FBS, low density (LD)/10% FBS, and low density/0.5% FBS probed with γ-smooth muscle (γ-SMA) and α-smooth muscle (α-SMA) isoactins on days (D) 1, 3, 5, 7, and 10 of culture. Quantitation values have been normalized to GAPDH expression. Relative expression of γ-smooth muscle (○) and α-smooth muscle (●) isoactins in cultured ISMCs plated on plastic at (B) low density/10% FBS, (C) high density/10% FBS, (E) high density/0.5% FBS, and (D) low density/0.5% FBS in isolated muscle strips (MS), immediately after enzyme dissociation (0), and at days (D) 1, 3, 5, 7, and 10 of culture.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

11. Fig. 3
(A) Northern blot analysis of cultured ISMCs plated at high density (HD)/10% FBS, high density/0.5% FBS, low density (LD)/10% FBS, and low density/0.5% FBS probed with γ-smooth muscle (γ-SMA) and α-smooth muscle (α-SMA) isoactins on days (D) 1, 3, 5, 7, and 10 of culture. Quantitation values have been normalized to GAPDH expression. Relative expression of γ-smooth muscle (○) and α-smooth muscle (●) isoactins in cultured ISMCs plated on plastic at (B) low density/10% FBS, (C) high density/10% FBS, (E) high density/0.5% FBS, and (D) low density/0.5% FBS in isolated muscle strips (MS), immediately after enzyme dissociation (0), and at days (D) 1, 3, 5, 7, and 10 of culture.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

12. Fig. 3
(A) Northern blot analysis of cultured ISMCs plated at high density (HD)/10% FBS, high density/0.5% FBS, low density (LD)/10% FBS, and low density/0.5% FBS probed with γ-smooth muscle (γ-SMA) and α-smooth muscle (α-SMA) isoactins on days (D) 1, 3, 5, 7, and 10 of culture. Quantitation values have been normalized to GAPDH expression. Relative expression of γ-smooth muscle (○) and α-smooth muscle (●) isoactins in cultured ISMCs plated on plastic at (B) low density/10% FBS, (C) high density/10% FBS, (E) high density/0.5% FBS, and (D) low density/0.5% FBS in isolated muscle strips (MS), immediately after enzyme dissociation (0), and at days (D) 1, 3, 5, 7, and 10 of culture.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

13. Fig. 3
(A) Northern blot analysis of cultured ISMCs plated at high density (HD)/10% FBS, high density/0.5% FBS, low density (LD)/10% FBS, and low density/0.5% FBS probed with γ-smooth muscle (γ-SMA) and α-smooth muscle (α-SMA) isoactins on days (D) 1, 3, 5, 7, and 10 of culture. Quantitation values have been normalized to GAPDH expression. Relative expression of γ-smooth muscle (○) and α-smooth muscle (●) isoactins in cultured ISMCs plated on plastic at (B) low density/10% FBS, (C) high density/10% FBS, (E) high density/0.5% FBS, and (D) low density/0.5% FBS in isolated muscle strips (MS), immediately after enzyme dissociation (0), and at days (D) 1, 3, 5, 7, and 10 of culture.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

14. Fig. 3
(A) Northern blot analysis of cultured ISMCs plated at high density (HD)/10% FBS, high density/0.5% FBS, low density (LD)/10% FBS, and low density/0.5% FBS probed with γ-smooth muscle (γ-SMA) and α-smooth muscle (α-SMA) isoactins on days (D) 1, 3, 5, 7, and 10 of culture. Quantitation values have been normalized to GAPDH expression. Relative expression of γ-smooth muscle (○) and α-smooth muscle (●) isoactins in cultured ISMCs plated on plastic at (B) low density/10% FBS, (C) high density/10% FBS, (E) high density/0.5% FBS, and (D) low density/0.5% FBS in isolated muscle strips (MS), immediately after enzyme dissociation (0), and at days (D) 1, 3, 5, 7, and 10 of culture.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

15. Fig. 4
(A) Northern blot analysis of cultured ISMCs plated at low density/10% FBS on fibronectin, laminin, and collagen type IV probed with γ-smooth muscle (γ-SMA) and α-smooth muscle (α-SMA) isoactins on days (D) 1, 3, 5, 7, and 10 of culture. Quantitation values have been normalized to GAPDH expression. Relative expression of γ-smooth muscle (●) and α-smooth muscle (○) isoactins in cultured ISMCs plated at low density/10% FBS on (B) plastic, (C) fibronectin, (D) laminin, and (E) collagen type IV in isolated muscle strips (MS), immediately after enzyme dissociation (0), and at days (D) 1, 3, 5, 7, and 10 of culture.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

16. Fig. 4
(A) Northern blot analysis of cultured ISMCs plated at low density/10% FBS on fibronectin, laminin, and collagen type IV probed with γ-smooth muscle (γ-SMA) and α-smooth muscle (α-SMA) isoactins on days (D) 1, 3, 5, 7, and 10 of culture. Quantitation values have been normalized to GAPDH expression. Relative expression of γ-smooth muscle (●) and α-smooth muscle (○) isoactins in cultured ISMCs plated at low density/10% FBS on (B) plastic, (C) fibronectin, (D) laminin, and (E) collagen type IV in isolated muscle strips (MS), immediately after enzyme dissociation (0), and at days (D) 1, 3, 5, 7, and 10 of culture.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

17. Fig. 4
(A) Northern blot analysis of cultured ISMCs plated at low density/10% FBS on fibronectin, laminin, and collagen type IV probed with γ-smooth muscle (γ-SMA) and α-smooth muscle (α-SMA) isoactins on days (D) 1, 3, 5, 7, and 10 of culture. Quantitation values have been normalized to GAPDH expression. Relative expression of γ-smooth muscle (●) and α-smooth muscle (○) isoactins in cultured ISMCs plated at low density/10% FBS on (B) plastic, (C) fibronectin, (D) laminin, and (E) collagen type IV in isolated muscle strips (MS), immediately after enzyme dissociation (0), and at days (D) 1, 3, 5, 7, and 10 of culture.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

18. Fig. 4
(A) Northern blot analysis of cultured ISMCs plated at low density/10% FBS on fibronectin, laminin, and collagen type IV probed with γ-smooth muscle (γ-SMA) and α-smooth muscle (α-SMA) isoactins on days (D) 1, 3, 5, 7, and 10 of culture. Quantitation values have been normalized to GAPDH expression. Relative expression of γ-smooth muscle (●) and α-smooth muscle (○) isoactins in cultured ISMCs plated at low density/10% FBS on (B) plastic, (C) fibronectin, (D) laminin, and (E) collagen type IV in isolated muscle strips (MS), immediately after enzyme dissociation (0), and at days (D) 1, 3, 5, 7, and 10 of culture.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

19. Fig. 4
(A) Northern blot analysis of cultured ISMCs plated at low density/10% FBS on fibronectin, laminin, and collagen type IV probed with γ-smooth muscle (γ-SMA) and α-smooth muscle (α-SMA) isoactins on days (D) 1, 3, 5, 7, and 10 of culture. Quantitation values have been normalized to GAPDH expression. Relative expression of γ-smooth muscle (●) and α-smooth muscle (○) isoactins in cultured ISMCs plated at low density/10% FBS on (B) plastic, (C) fibronectin, (D) laminin, and (E) collagen type IV in isolated muscle strips (MS), immediately after enzyme dissociation (0), and at days (D) 1, 3, 5, 7, and 10 of culture.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

20. Fig. 5
(A) Proposed model of rat gastrointestinal smooth muscle development in vivo at embryonic days 9 (E9), 11 (E11), 15 (E15), and adult showing mucosa (MU), mesenchyme (ME), serosa (SE), primitive circular smooth muscle layer (PCM), primitive longitudinal smooth muscle layer (PLM), primitive muscularis mucosa (PMM), circular smooth muscle layer (CM), longitudinal smooth muscle layer (LM), and muscularis mucosa (MM). (B) Cellular phenotypes principally observed at these developmental time points with the corresponding phenotypes observed at day 1 (D1) and day 10 (D10) of culture marked. Gene products known to be expressed in the various smooth muscle cell phenotypes are highlighted in boxes.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions