Induced pluripotent stem cell-derived enteric neural crest cells repopulate human aganglionic tissue-engineered intestine to form key components of the enteric nervous system

David F Chang; Samuel M Zuber; Elizabeth A Gilliam; Laura-Marie A Nucho; Gabriel Levin; Fengnan Wang; Anthony I Squillaro; Sha Huang; Jason R Spence; Tracy C Grikscheit

doi:10.1177/2041731420905701

Induced pluripotent stem cell-derived enteric neural crest cells repopulate human aganglionic tissue-engineered intestine to form key components of the enteric nervous system

J Tissue Eng. 2020 Apr 27:11:2041731420905701. doi: 10.1177/2041731420905701. eCollection 2020 Jan-Dec.

Authors

David F Chang¹, Samuel M Zuber¹, Elizabeth A Gilliam¹, Laura-Marie A Nucho¹, Gabriel Levin¹, Fengnan Wang¹, Anthony I Squillaro¹, Sha Huang^{2

3}, Jason R Spence^{2

3

4}, Tracy C Grikscheit^{1

5

6}

Affiliations

¹ Developmental Biology and Regenerative Medicine Program, The Saban Research Institute, Children's Hospital Los Angeles, Los Angeles, CA, USA.
² Department of Internal Medicine, Medical School, University of Michigan, Ann Arbor, MI, USA.
³ Department of Cell & Developmental Biology, Medical School, University of Michigan, Ann Arbor, MI, USA.
⁴ Program of Cellular & Molecular Biology, Medical School, University of Michigan, Ann Arbor, MI, USA.
⁵ Department of Surgery, Division of Pediatric Surgery, Children's Hospital Los Angeles, Los Angeles, CA, USA.
⁶ Keck Medical School, University of Southern California, Los Angeles, CA, USA.

Abstract

Models for enteric neuropathies, in which intestinal nerves are absent or injured, are required to evaluate possible cell therapies. However, existing options, including transgenic mice, are variable and fragile. Here immunocompromised mice were implanted with human pluripotent stem cell-derived tissue-engineered small intestine 10 weeks prior to a second survival surgery in which enteric nervous system precursor cells, or saline controls, were injected into the human intestinal organoid-derived tissue-engineered small intestine and analyzed 4 weeks later. Human intestinal organoid-derived tissue-engineered small intestine implants injected with saline as controls illustrated formation of intestinal epithelium and mesenchyme without an enteric nervous system. Second surgical introduction of human pluripotent stem cell-generated enteric nervous system precursors into developing human intestinal organoid-derived tissue-engineered small intestine implants resulted in proliferative migratory neuronal and glial cells, including multiple neuronal subtypes, and demonstrated function in contractility assays.

Keywords: Hirschsprung disease; Tissue-engineered small intestine; enteric nervous system; human intestinal organoids.