578 | Nature | Vol 585 | 24 September 2020
Article
experiments reveal that our bioengineered organoids show notable
regenerative potential.
Modelling long-term parasite infection
Finally, we investigated whether mini-gut tubes could be used to model
long-term infection caused by Cryptosporidium parvum, an obligate par-
asite that results in life-threatening diarrhoea in immunocompromised
adult hosts and in infants^17. Research on the pathophysiology of C. par-
vum has been hindered by the lack of long-term, primary-cell-derived
in vitro culture systems. Conventional 3D organoids can be used to
model C. parvum infection^18 , but the luminal inaccessibility and inability
of the system to support long-term host–microorganism co-cultures
limit the applicability of this system.
We infected mini-gut tubes with C. parvum by loading a suspension of
oocysts into the inlet reservoir of the microchip (Extended Data Fig. 9a).
Live-cell microscopy demonstrated that the tubular organoids support
the completion of the life cycle and long-term growth of C. parvum
without compromising tissue integrity (Extended Data Fig. 9b, Sup-
plementary Video 9). The identity of each asexual and sexual stage was
confirmed using immunofluorescence assays (Fig. 3d) and TEM imaging
on mini-gut cross-sections (Fig. 3f, Extended Data Fig. 9c, d). By infect-
ing mini-guts with freshly isolated sporozoites and analysing the lumi-
nal content every day, we observed successive rounds of production of
newly formed oocysts for at least four weeks (Extended Data Fig. 9e).
Gene set enrichment analysis of infected samples showed a signifi-
cant enrichment of interferon-α response genes, as well as changes in
metabolism (Extended Data Fig. 9f ). An analysis by cell type showed that
the interferon response was not limited to one specific population, but
was instead distributed across all cell types, even though the response
genes showed some cell-type specificities (Extended Data Fig. 9g; see
also Source Data). Altogether, these results show that these bioen-
gineered organoids, similar to primary-stem-cell-derived intestinal
monolayers comprising feeder cells^19 , are ideally suited for mechanistic
host–microorganism interaction and long-term infection studies.
By combining bioengineering with the self-organization properties of
stem cells, here we have generated open, tubular ‘organoids-on-a-chip’
that exhibit exceptional cell-type diversity, tissue architecture and func-
tion. The introduction of a microchip-based perfusion system made it
possible to efficiently remove shed cells from the lumen and expose it
to parasites or medium additives. Previous efforts to model intestinal
epithelia through micro-engineering or tissue engineering^7 ,^8 ,^19 –^25 have
successfully addressed the problem of lumen accessibility, but it has
not yet been possible in these systems to capture the diversity of cell
types and the patterning that are found in vivo or in classical organoids.
Moreover, existing approaches are based on polarized cell monolay-
ers grown on two-dimensional permeable polymer membranes^26 ,^27 ,
which may preclude the modelling of complex 3D multi-tissue interac-
tions. The biomimetic 3D extracellular matrix that surrounds the pat-
terned intestinal epithelium in our system can be readily colonized with
non-epithelial cell types (Extended Data Fig. 10a) such as endothelial
cells (Extended Data Fig. 10b), immune cells (Extended Data Fig. 10c, d)
and myofibroblasts (Extended Data Fig. 10e, f ). These supportive cell
types were found to communicate with intestinal epithelial cells; for
example, macrophages were observed to undergo morphological
changes and ingest particles excreted from the epithelium (Extended
Data Fig. 10c, Supplementary Video 10). The functional integration of
an immune axis in bioengineered, homeostatic organoids opens up
new perspectives for disease modelling. We anticipate that by adjust-
ing specific characteristics of the hydrogel scaffolds (for example,
their composition, geometry, size, stiffness and signalling inputs), the
approach we describe here could be applied to other organoid-forming
stem cells—including those derived from other organs, such as the
lung, liver or pancreas, and derived from patient biopsies (Extended
Data Fig. 2). The readily accessible 3D tissue anatomy of our model,
which captures the development of stem cells in a highly tractable
experimental framework, will answer questions that have so far been
difficult to address, and may have substantial potential for drug dis-
covery, diagnostics and regenerative medicine.Online content
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maries, source data, extended data, supplementary information,
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