micropatterned systems rely on the supplementation with Matrigel
to provide apicobasal polarization cues to allow cells to attain a 3D
architecture.
2 Material
Prepare all solutions using ultrapure water and analytical grade
reagents. Prepare and store all reagents as specified by the manufacturer. For cell culture purposes, keep sterile conditions. Carefully
follow the waste disposal regulations.
2.1 Fabrication
of Flat Micropatterns
1. Photolithographed silicon wafer with the desired
microfeatures.
2. Trichloro(1H,1H,2H,2H-perfluorooctyl)silane.
3. Oven (should reach 80
C).
4. Desiccator.
5. Vacuum pump.
6. Hood.
7. UV/O 3 cleaner.
8. Spin coater.
9. Borosilicate glass Petri dishes.
10. 18 mm coverslips.
11. 6-Well plates.
12. 1.5 ml tubes.
13. PDMS (Sylgard 184).
14. Laminin from Engelbreth-Holm-Swarm murine sarcoma basement membrane.
15. Human plasma fibronectin purified protein or RhodamineFibronectin.
16. Pluronic F127.
17. MilliQ water.
18. Phosphate-buffered saline (PBS).
19. Compressed air duster.
20. Sharp scalpel and single edge razor, curved forceps.
2.2 Fabrication
of the 3D Microwells
1. Photolithographed silicon wafer or counter-mold with the
desired microfeatures.
2. Oven (should reach 80
C).
3. Desiccator.
4. Vacuum pump.
Micropatterned Tubes for Epithelial Analyses
229
to provide apicobasal polarization cues to allow cells to attain a 3D
architecture.
2 Material
Prepare all solutions using ultrapure water and analytical grade
reagents. Prepare and store all reagents as specified by the manufacturer. For cell culture purposes, keep sterile conditions. Carefully
follow the waste disposal regulations.
2.1 Fabrication
of Flat Micropatterns
1. Photolithographed silicon wafer with the desired
microfeatures.
2. Trichloro(1H,1H,2H,2H-perfluorooctyl)silane.
3. Oven (should reach 80
C).
4. Desiccator.
5. Vacuum pump.
6. Hood.
7. UV/O 3 cleaner.
8. Spin coater.
9. Borosilicate glass Petri dishes.
10. 18 mm coverslips.
11. 6-Well plates.
12. 1.5 ml tubes.
13. PDMS (Sylgard 184).
14. Laminin from Engelbreth-Holm-Swarm murine sarcoma basement membrane.
15. Human plasma fibronectin purified protein or RhodamineFibronectin.
16. Pluronic F127.
17. MilliQ water.
18. Phosphate-buffered saline (PBS).
19. Compressed air duster.
20. Sharp scalpel and single edge razor, curved forceps.
2.2 Fabrication
of the 3D Microwells
1. Photolithographed silicon wafer or counter-mold with the
desired microfeatures.
2. Oven (should reach 80
C).
3. Desiccator.
4. Vacuum pump.
Micropatterned Tubes for Epithelial Analyses
229