In vitro Dynamic Model of a Catheterized Bladder and Biofilm Assay

Materials and Reagents

1. NaCl (Carl Roth, catalog number: 9265.1 )
   
2. 70% ethanol
   
3. dH₂O
   
4. NaCl (Carl Roth, catalog number: 9265.1)
   
5. Tryptone (Carl Roth, catalog number: 8952.3 )
   
6. Yeast extract (Carl Roth, catalog number: 2362.2 )
   
7. Agar-agar (Carl Roth, catalog number: 5210.2 )
   
8. Na₂SO₄ (Carl Roth, catalog number: 8560.1 )
   
9. CaCl₂^.2H₂O (Carl Roth, catalog number: CN93.1 )
   
10. MgCl₂^.6H₂O (Carl Roth, catalog number: 2189.1 )
    
11. NaCl (Carl Roth, catalog number: 9265.1)
    
12. Na₃C₆H₅O₇^.2H₂O (Carl Roth, catalog number: 3580.1 )
    
13. (COONa)₂ (Carl Roth, catalog number: 4267.1 )
    
14. KH₂PO₄ (Carl Roth, catalog number: 3904.1 )
    
15. KCl (Carl Roth, catalog number: 6781.1 )
    
16. NH₄Cl (Carl Roth, catalog number: K298.2 )
    
17. Urea (Carl Roth, catalog number: X999.3 )
    
18. Tryptic soya broth/CASO medium (Carl Roth, catalog number: X938.1 )
    
19. Gelatine (Carl Roth, catalog number: 4582.2 )
    
20. NaOH (Carl Roth, catalog number: 6771.1 )
    
21. LB medium (see Recipes)
    
22. LB agar (see Recipes)
    
23. Artificial or human urine (see Recipes)
    

Equipment

1. Peristaltic pump for urine filtration (e.g. Heidolph Instruments GmbH, model: Pump drive 5006 )
   
2. 10 L polycarbonate carboy for urine reservoir (e.g. Thermo Fisher Scientific, Nalgene^®, 10 L PC with polypropylene cap with three inlets)
   
3. SARTORIUS P Midi Cap filter cartridge (SARTORIUS, catalog number: 5235307H9 )
   
4. Peristaltic multi-channel pump for urine feed (Watson-Marlow Pumps Group, model: 205U )
   
5. Borosilicate Glass bladder models (Figure 1) (Georg Becker Laboreinrichtungen GmbH & Co KG, www.laborbecker.at)
   
   
   Figure 1. Glass bladder model scheme and dimensions. Schematic representation including length and inner diameter (ID) indications A and custom-made model B. Relevant parts are indicated: Artificial bladder compartment (1), temperature control compartment (2), connection for incoming (3) and outgoing (4) temperature control water circuit, artificial urethra for catheter insertion (5).
   
   
6. Water bath with connections for external water circle (e.g. LAUDA, model: E10 )
   
7. Tripod or other holders for mounting of glass bladder models (e.g. Bochem Instrumente GmbH, Portable table frame TG, catalog number: TG500 )
   
8. Silicone stopper (35 mm) with 6 mm decentralized hole
   
9. Glass tube (15 cm, 6 mm outer diameter = OD)
   Note: We use cut 1 ml glass pipettes (BRAND).
   
10. Silicone tubings: 2 m (ID 3 mm, OD 5 mm)
    
11. Silicone tubings: 1 m (ID 9 mm)
    
12. Silicone tubings: 1 m (ID 5-6 mm)
    
13. Tygon tubings (ID 8 mm) (VWR International, catalog number: 228-1294 )
    
14. Tube Connectors (PP, 3-5mm - 6-10 mm) (Bartelt, catalog number: 37.559 )
    
15. T-connectors (PP) for tubings (ID 3 mm) (Carl Roth, catalog number: E763.1 )
    
16. Urinary Catheter (all silicone Ch14) (BARD, catalog number: 153509 )
    
17. 10 ml syringe
    
18. Drainage bags (e.g. SARSTEDT AG, catalog number: 74.5220.005 )
    
19. Clamps (e.g. VWR International, catalog number: 229-0072 )
    
20. Ultrasonic cleaner (BANDELIN electronic, model: Sonorex RK100H )
    
21. Vortex (e.g. VWR International, catalog number: 444-0206 )
    
22. Recommended (air vents as breathing barrier for urine reservoir flask) (e.g. VWR International, catalog number: 28144-160 )
    

Procedure