An Ex Vivo Gut Organ Culture System to Study Host-Microbiota Interactions
Published: September 29, 2023
Abstract
Source: Azriel, S., et al. An Intestinal Gut Organ Culture System for Analyzing Host-Microbiota Interactions. J. Vis. Exp. (2021).
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…
開示
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) | Thermo Fisher Scientific | 12440061 | |
| HEPES Buffer (1M) | Thermo Fisher Scientific | 15630056 | |
| Knock-Out Serum | Thermo Fisher Scientific | 10828028 | |
| N2 Supplement (100x) | Thermo Fisher Scientific | A1370701 | |
| Non-Essential Amino Acid (100x) | Thermo Fisher Scientific | 11140035 | |
| Surgical Tools | |||
| Large Scissors | Aseltech | 11-00-10 | |
| Sharp Forceps | F.S.T | 11297-10 | |
| Silk-Braided Surgical Thread | SMI | 8010G | |
| Straight Scissors | F.S.T | 14091-09 | |
| Thin Forceps | F.S.T | 11051-10 | |
| Organ System | |||
| 0.1 µm Filter | Life Gene | ||
| 0.22 µm Filter | Life Gene | ||
| 5 mL Luer-Lock Syringe | B-D | 309649 | |
| Greenough Stereo Microscope | ZEISS | Stemi 305 | |
| Recirculating Precision Air Heater "CUBE" | CUBE-2-LIS | ||
| Syringe Pump | New Era Pump Systems Inc | nep-ne-1600-em |
