Blood, Vol. 114, Issue 2, 237-247, July 9, 2009

Endothelial cell lumen and vascular guidance tunnel formation requires MT1-MMP–dependent proteolysis in 3-dimensional collagen matrices
Blood Stratman et al. 114: 237

Supplemental materials for: Stratman et al

Files in this Data Supplement:

• Video 10. MT1-MMP siRNA treated ECs show marked blockade of motility as well as inhibited EC lumen and vascular guidance tunnel formation in 3D collagen matrices (MOV, 1.66 MB) -
  EC cultures were established with nuclear GFP-labeled ECs in 3D collagen matrices (after treatment with MT1-MMP siRNA) and were imaged by time-lapse fluorescence and light microscopy over a 24 hr period. Frames were collected every 10 minutes, were overlaid, and are shown at 8 frames/sec.
  
  
• Video 11. EC tube remodeling and motility within vascular guidance tunnels present in 3D collagen matrices (MOV, 3.91 MB) -
  EC cultures were established for 48 hr and then were imaged by time-lapse microscopy over a 29 hr period. Frames were collected every 10 minutes and are shown at 6 frames/sec.
  
  
• Video 12. EC motility within vascular guidance tunnels present in 3D collagen matrices in the presence of the MMP inhibitor, GM6001 (MOV, 2.14 MB) -
  EC cultures were established for 48 hr in the absence of proteinase inhibitors and then were imaged by time-lapse microscopy over a 48 hr period in the presence of GM6001 at 5 µM (i.e. added after 48 hrs of culture). Frames were collected every 10 minutes and are shown at 5 frames/sec.
  
  
• Video 13. EC tube regrowth within vascular guidance tunnels following thrombin-induced collapse and reversal with hirudin (MOV, 1.50 MB) -
  EC cultures were established for 48 hr and then were treated with 10 µg/ml of thrombin for 30 minutes followed by reversal with hirudin. The movie extends from 48 to 56 hr. Frames were collected every 10 minutes and are shown at 2 frames/sec.
  
  
• Video 14. Control EC motility within vascular guidance tunnels present in 3D collagen matrices (MOV, 2.52 MB) -
  EC cultures were established for 48 hr and then were imaged by time-lapse microscopy over a 24 hr period. Frames were collected every 10 minutes and are shown at 10 frames/sec. Please note that the networks of EC-lined tube structures remain intact and modestly expand over this time course.
  
  
• Video 15. Anti-α2 integrin antibodies alter vascular tube remodeling within vascular guidance tunnels present in 3D collagen matrices (MOV, 1.56 MB) -
  EC cultures were established for 48 hr and then were imaged by time-lapse microscopy over a 24 hr period in the presence of anti-α2 integrin antibodies added at 20 µg/ml (i.e. added after 48 hrs of culture). Frames were collected every 10 minutes and are shown at 10 frames/sec.
  
  
• Video 16. Anti-β1 integrin antibodies alter vascular tube remodeling within vascular guidance tunnels present in 3D collagen matrices (MOV, 1.87 MB) -
  EC cultures were established for 48 hr and then were imaged by time-lapse microscopy over a 24 hr period in the presence of anti-β1 integrin antibodies added at 20 µg/ml (i.e. added after 48 hrs of culture). Frames were collected every 10 minutes and are shown at 10 frames/sec.
  
  
• Video 17. Anti-α5 integrin antibodies have no influence on vascular tube remodeling within vascular guidance tunnels present in 3D collagen matrices (MOV, 1.8 MB) -
  EC cultures were established for 48 hr and then were imaged by time-lapse microscopy over a 24 hr period in the presence of anti-α5 integrin antibodies added at 20 µg/ml (i.e. added after 48 hrs of culture). Frames were collected every 10 minutes and are shown at 10 frames/sec.