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Generating 3D human intestinal organoids with an enteric nervous system

Workman, Michael J.
http://rave.ohiolink.edu/etdc/view?acc_num=ucin1416570664

Abstract Details

2014, MS, University of Cincinnati, Medicine: Molecular and Developmental Biology.
The ENS arises from neural crest cells (NCCs) to form the largest division of the vertebrate peripheral nervous system and is essential for gastrointestinal (GI) motility, secretion, epithelial permeability and fluid exchange. It is estimated that nearly 30% of the Western population is affected by what are presumed to be functional disorders of the ENS and GI tract. Despite this fact, no in vitro human model exists to study the ENS and very few drugs are available to target the deficiencies associated with enteric neuropathies. Our lab has recently developed a 3D human intestinal organoid (HIO) culture system based on the directed differentiation of pluripotent stem cells (PSCs); however, these do not contain an ENS. Here we show the generation organoids containing functional neural networks resembling an ENS by combining early stage HIOs with NCCs. We have successfully established a protocol to generate NCCs from human PSCs and have identified that retinoic acid signaling is capable of directing cranial-like NCCs to a more vagal-like fate, which represent the main progenitors of the ENS. When differentiated in vitro, NCCs form neurons, glia, chondrocytes, and osteocytes. Additionally, engraftment of NCCs into chick embryos show that PSC-derived NCCs can migrate along predicted paths and be found associated with the developing chick foregut. When combined with early stage HIOs, NCCs display robust neural engraftment and association with intestinal tissue reminiscent of early gut development. In vivo engraftment of HIOs+NCCs into the kidney capsule of immunodeficient mice promotes organoid maturation and results in complex organoids displaying vilified, columnar epithelium surrounded by layers of smooth muscle containing a closely associated neural plexus. HIOs+NCCs develop a large diversity of neural cell types which show rhythmic Ca2+ flux and responsiveness to KCl. Further investigation into neuronal function and impact of ENS on GI development is needed; however, these organoids offer a novel platform for modeling GI motility disorders and screening genetic and chemical factors that affect motility and secretion.
James Wells, Ph.D. (Committee Chair)
Samantha Brugmann, Ph.D. (Committee Member)
Aaron Zorn, Ph.D. (Committee Member)
60 p.
Developmental Biology
enteric nervous system; intestinal organoids; neural crest; human pluripotent stem cells; tissue engineering

Recommended Citations

Citations

  • Workman, M. J. (2014). Generating 3D human intestinal organoids with an enteric nervous system [Master's thesis, University of Cincinnati]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1416570664

    APA Style (7th edition)

  • Workman, Michael. Generating 3D human intestinal organoids with an enteric nervous system. 2014. University of Cincinnati, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ucin1416570664.

    MLA Style (8th edition)

  • Workman, Michael. "Generating 3D human intestinal organoids with an enteric nervous system." Master's thesis, University of Cincinnati, 2014. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1416570664

    Chicago Manual of Style (17th edition)

ucin1416570664
595
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This open access ETD is published by University of Cincinnati and OhioLINK.