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

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記事を引用
An Ex Vivo Gut Organ Culture System to Study Host-Microbiota Interactions. J. Vis. Exp. (Pending Publication), e21617, doi: (2023).

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