5. Hood.
6. UV/O 3 cleaner.
7. Spin coater.
8. Borosilicate glass Petri dishes.
9. 60 mm Bacteriological Petri dishes.
10. 18 mm coverslips.
11. 6-Well plates.
12. PDMS (Sylgard 184).
13. Rhodamine-fibronectin.
14. MilliQ water.
15. Phosphate-buffered saline (PBS).
16. Sharp scalpel and single edge razor, curved forceps.
17. Fluorescence microscope.
2.3 Culture of MDCK
Cells as 3D Tubes
1. Tissue-culture hood.
2. Flat patterned surface (either CYTOO chip or μCPrinted chip)
or 3D substrates with microchannels.
3. Centrifuge.
4. MDCK II cells.
5. Ice bucket.
6. MEM medium supplemented with 10% fetal bovine serum
(FBS), 2 mM L-glutamine, 100 U/ml penicillin, and
100 mg/ml streptomycin for cell maintenance.
7. 0.25% (w/v) trypsin/1 mM EDTA.
8. Tissue-culture dishes.
9. Hemocytometer.
10. 15 ml conical tube.
11. Matrigel.
2.4 Culture of Mice
Mammary Gland
Organoids on Line
Microchannels
1. Tissue-culture hood.
2. ECM-coated PDMS substrates with microwells.
3. Centrifuge suitable for plates.
4. Isolated mice mammary gland organoids.
5. DMEM/F12.
6. Fetal bovine serum.
7. Bovine serum albumin.
8. Ice bucket.
9. DispaseII.
10. DNaseI.
230
Minerva Bosch-Fortea and Fernando Martı ´n-Belmonte
6. UV/O 3 cleaner.
7. Spin coater.
8. Borosilicate glass Petri dishes.
9. 60 mm Bacteriological Petri dishes.
10. 18 mm coverslips.
11. 6-Well plates.
12. PDMS (Sylgard 184).
13. Rhodamine-fibronectin.
14. MilliQ water.
15. Phosphate-buffered saline (PBS).
16. Sharp scalpel and single edge razor, curved forceps.
17. Fluorescence microscope.
2.3 Culture of MDCK
Cells as 3D Tubes
1. Tissue-culture hood.
2. Flat patterned surface (either CYTOO chip or μCPrinted chip)
or 3D substrates with microchannels.
3. Centrifuge.
4. MDCK II cells.
5. Ice bucket.
6. MEM medium supplemented with 10% fetal bovine serum
(FBS), 2 mM L-glutamine, 100 U/ml penicillin, and
100 mg/ml streptomycin for cell maintenance.
7. 0.25% (w/v) trypsin/1 mM EDTA.
8. Tissue-culture dishes.
9. Hemocytometer.
10. 15 ml conical tube.
11. Matrigel.
2.4 Culture of Mice
Mammary Gland
Organoids on Line
Microchannels
1. Tissue-culture hood.
2. ECM-coated PDMS substrates with microwells.
3. Centrifuge suitable for plates.
4. Isolated mice mammary gland organoids.
5. DMEM/F12.
6. Fetal bovine serum.
7. Bovine serum albumin.
8. Ice bucket.
9. DispaseII.
10. DNaseI.
230
Minerva Bosch-Fortea and Fernando Martı ´n-Belmonte