Culture and differentiation of rabbit intestinal organoids and organoid-derived cell monolayers

Egi Kardia, Michael Frese, Elena Smertina, Tanja Strive, Xi-Lei Zeng, Mary Estes, Robyn N Hall

Research output: Contribution to journalArticlepeer-review

3 Downloads (Pure)

Abstract

Organoids emulate many aspects of their parental tissue and are therefore used to study pathogen-host interactions and other complex biological processes. Here, we report a robust protocol for the isolation, maintenance and differentiation of rabbit small intestinal organoids and organoid-derived cell monolayers. Our rabbit intestinal spheroid and monolayer cultures grew most efficiently in L-WRN-conditioned medium that contained Wnt, R-spondin and Noggin, and that had been supplemented with ROCK and TGF-β inhibitors. Organoid and monolayer differentiation was initiated by reducing the concentration of the L-WRN-conditioned medium and by adding ROCK and Notch signalling inhibitors. Immunofluorescence staining and RT-qPCR demonstrated that our organoids contained enterocytes, enteroendocrine cells, goblet cells and Paneth cells. Finally, we infected rabbit organoids with Rabbit calicivirus Australia-1, an enterotropic lagovirus that-like many other caliciviruses-does not grow in conventional cell culture. Despite testing various conditions for inoculation, we did not detect any evidence of virus replication, suggesting either that our organoids do not contain suitable host cell types or that additional co-factors are required for a productive infection of rabbit organoids with Rabbit calicivirus Australia-1.

Original languageEnglish
Article number5401
Pages (from-to)1-12
Number of pages12
JournalScientific Reports
Volume11
Issue number1
DOIs
Publication statusPublished - 8 Mar 2021

Fingerprint

Dive into the research topics of 'Culture and differentiation of rabbit intestinal organoids and organoid-derived cell monolayers'. Together they form a unique fingerprint.

Cite this