Perfusion and control of microbiome in lumen of intestinal organoids

Intestinal organoids have found application in modeling intestinal function and infection, but is it feasible to replicate gastrointestinal motility and microbiome stability with organoids? One of the limitations is to access their lumens and modify their contents. A challenge that Ginga et al. at the Georgia Institute of Technology address, using a perfusion system, modeling host-microbe interaction in human intestinal organoids.

The authors were able to perfuse E. coli into the intestinal organoids. In a static lumen resulted the epithelium structural integrity was lost, whereas the perfused organoids showed approximately two times larger epithelial thickness, improved structural integrity and epithelial polarization. Moreover, luminal perfusion enhanced the organoids functional maturation shown by higher expression of DPPIV (a digestive enzyme).

DigesTable of the paper Ginga, N.J.; Slyman, R.; Kim, G.-A.; Parigoris, E.; Huang, S.; Yadagiri, V.K.; Young, V.B.; Spence, J.R.; Takayama, S. Perfusion System for Modification of Luminal Contents of Human Intestinal Organoids and Realtime Imaging Analysis of Microbial Populations. Micromachines 202213, 131. https://doi.org/10.3390/mi13010131. This paper is reproduced under https://creativecommons.org/licenses/by/4.0/. The image of the chip was edited for better clarity, data in the table and text were compiled and interpreted by AZAR Innovations.

The authors used a holder to keep the organoid in place and two glass capillary tubes to perfuse pluripotent stem-cell-derived intestinal organoids.
3D printing was used to make a mold for the PDMS holder
– Sterilization
: not mentioned!
– Organoid incorporation: the holder is open from the top, organoids and Matrigel were placed from the top
Perfusion/refreshing: injecting and withdrawing the fluid by two glass capillary tubes using a
pressure-based pump
– Treatment: no treatment

On-chip read-outs: real-time microscopy
– Off-chip read-outs:
off-chip imaging (IHC)

Strong points :
+ Control of luminal content in time using a
pressure-based pump
+ Multi-focal imaging and analysis of the bacterial content
+ Robust

Nothing is perfect, some improvement points:
Perfusion is short with a high chance of contamination
– Limited perfusion-induced cyclic strain compared to some intestine-on-chip devices
– Limited throughput
– No negative control without E- coli
– Comparison between imaging during perfusion and pre-/post-perfusion is not feasible
– Co-culture with autologous cell types for personalized medicine applications
would be awesome!

Conclusion and outlook
This paper was an effort to access the lumen of intestinal organoids and add microbiome to study host-microbe interaction. The perfusion technology of this study can also be used for other infection models, such as for lung infection. 

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