Perfusion System for Modification of Luminal Contents of Human Intestinal Organoids and Realtime Imaging Analysis of Microbial Populations

Intestinal organoids are 3D cell structures that replicate some aspects of organ function and are organized with a polarized epithelium facing a central lumen. To enable more applications, new technologies are needed to access the luminal cavity and apical cell surface of organoids. We developed a p...

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Bibliographic Details
Published in:	Micromachines (Basel) Vol. 13; no. 1; p. 131
Main Authors:	Ginga, Nicholas J, Slyman, Raleigh, Kim, Ge-Ah, Parigoris, Eric, Huang, Sha, Yadagiri, Veda K, Young, Vincent B, Spence, Jason R, Takayama, Shuichi
Format:	Journal Article
Language:	English
Published:	Switzerland MDPI AG 14-01-2022 MDPI
Subjects:	Bacteria Blood vessels Capillary pressure Cell culture Design Dextrans E coli Enumeration Fluorescence intestine microfluidics Microorganisms New technology organoid perfusion Physiology pump Real time Time measurement United States > US intestine perfusion bacteria fluorescence pump image analysis injection microfluidics PDMS organoid
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Summary:	Intestinal organoids are 3D cell structures that replicate some aspects of organ function and are organized with a polarized epithelium facing a central lumen. To enable more applications, new technologies are needed to access the luminal cavity and apical cell surface of organoids. We developed a perfusion system utilizing a double-barrel glass capillary with a pressure-based pump to access and modify the luminal contents of a human intestinal organoid for extended periods of time while applying cyclic cellular strain. Cyclic injection and withdrawal of fluorescent FITC-Dextran coupled with real-time measurement of fluorescence intensity showed discrete changes of intensity correlating with perfusion cycles. The perfusion system was also used to modify the lumen of organoids injected with GFP-expressing . Due to the low concentration and fluorescence of the , a novel imaging analysis method utilizing bacteria enumeration and image flattening was developed to monitor within the organoid. Collectively, this work shows that a double-barrel perfusion system provides constant luminal access and allows regulation of luminal contents and luminal mixing.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 These authors contributed equally to this work.
ISSN:	2072-666X 2072-666X
DOI:	10.3390/mi13010131