Bioengineered In Vitro 3D Human Enteric Nervous System.
Manousiouthakis, Eleana.
2019
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The Gut-Brain axis is
lauded as an avenue to understand and remedy many neurological and gastrointestinal
disorders, with a focus on the gut microbiome in the years following the NIH human
microbiome project [1-4]. The field was sparked in the early 2000s by the discovery that
germ free mice showed enhanced response to stress by way of the
hypothalamic-pituitary-adrenal (HPA) reaction, and ... read morethis response could be ameliorated
using certain pathogen free species [5]. The gut-brain axis is thought to include the
central nervous system (CNS), the autonomic nervous system (ANS), the HPA, and the
enteric nervous system (ENS) [6]. The ENS is the first neural system that gut content
communicates with and thus represents a large target for bacterial metabolites. These
microbial communities have an intimate relationship with host tissue and have been
implicated in mucosal immune activation, reference memory, and anxiety-like behavior in
mouse models [6, 7]. Much of the research being performed with respect to the ENS and
the gut-brain axis is in animal models/tissues or in in vitro systems that currently do
not have microbial capabilities [8, 9]. Our group has previously developed a silk-based
scaffold with the capability of housing oxygen sensitive pathogenic and commensal
microbes [10-12]. The goal of this work was to develop a 3D human in vitro intestinal
tissue model to emulate the intricate cell interactions of the human enteric nervous
system and the epithelial layer utilizing the previously developed model in order to
better understand complex human conditions and microbial interactions that cannot be
studied in vivo. Here we describe a 3D innervated tissue model of the human intestine
consisting of human induced neural stem cells (hiNSCs) differentiated into relevant ENS
neural cell types, enterocyte-like (Caco-2) and goblet-like (HT29-MTX) cells forming the
intestinal epithelial layer, and intestinal myofibroblasts (IMF) as the stromal layer.
In vitro enteric nervous system (ivENS) cultures supported survival and function of the
various cell biomarkers (nNOS, TAC1, NK1, MUC2, Muc5AC, and TuJ1), with mucosal and
neural transcription factors evident over 5 weeks. Next, ivENS were inoculated using
microbial metabolites, taurine (T) and kynurenate (K), prior to treatment with
lipopolysaccharides (LPS) from E. coli. Results from these various treatment conditions
indicated that metabolites contribute a potential protective effect against the
pro-inflammatory response of cytokine monocyte chemoattractant protein‐1 (MCP-1)
production. This innervated tissue system offers a new tool to use to help in
understanding neural circuits controlling the human intestine and associated
communication networks.
Thesis (Ph.D.)--Tufts University, 2019.
Submitted to the Dept. of Biomedical Engineering.
Advisor: David Kaplan.
Committee: Kyongbum Lee, Kara Gross Margolis, and Madeleine Oudin.
Keywords: Biomedical engineering, Neurosciences, and Bioengineering.read less - ID:
- 8336hf083
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