In this video, we describe the methodology for setting up an ex vivo gut organ culture system to study the interactions between mouse intestinal tissue fragments and specific gut bacteria. The device consists of two pairs of input-output ports, the first connecting the intestinal tissue lumen and supplying the specific bacterial suspension into it, and the second continuously supplying media to the device's wells to maintain ex vivo tissue viability.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Experiment preparation Fabrication of the gut organ culture device (3 days) Using a 3D printer, print the reusable plastic molds for the organ culture device (the device has 6 wells, with 24 small and large holes, and for the device cover lid). NOTE: These plastic molds may be used f…
Declarações
The authors have nothing to disclose.
Materials
Device
18-Gauge Blunt Needle
Mcmaster
75165a754
22-Gauge Blunt Needle
Mcmaster
75165a758
All Purpose Adhesive Sealant 100% Silicone
DAP
688
Cubic Vacuum Desiccator VDC-21+ 2 Shelves
AAAD4021
Glass Slide 1 mm Thick
Corning
2947-75X50
Mini Incubator im-10
AAH24315K
MPC 301E Vacuum PUMP
VI-412711
Plastic Quick Turn Tube Coupling Plugs
Mcmaster
51525k121
Plastic Quick Turn Tube Coupling Sockets
Mcmaster
52525k211
Sylgard 184 Silicone Elastomer
Dow
Polydimethylsiloxane, PDMS
Tubing
Mcmaster
6516t11
Zortrax M200
Zortrax
Zortrax Z-SUITE, Autodesk Fusion 360
Zortrax M200 Materials: z-ultrat
Zortrax
Medium
B27 Supplement (50x), Serum-Free T
Thermo Fisher Scientific
17504044
Iscove's Modified Dulbecco's Medium with Phenol Red (1x)