Authors: Akaljot Singh, Holly M. Poling, Praneet Chaturvedi, Praneet Chaturvedi, Konrad Thorner, Nambirajan Sundaram, Nambirajan Sundaram, Charlie J. Childs, Heather McCauley, Garret W. Fisher, Nicole E. Brown, Jason R. Spence, James M. Wells, Michael A. Helmrath

Abstract: The in vitro differentiation of pluripotent stem cells into human intestinal organoids (HIOs) has served as a powerful means for creating cellularly complex three-dimensional intestinal structures. Due to their diverse cell populations, transplantation and engraftment are supported with this system and allow for enhanced development and maturation. Using an immunocompromised host as a bioreactor, transplanted HIOs form into fully laminated structures including a diverse epithelium with crypt-villus architecture and smooth muscle layers which resemble native human intestine. While the endpoint of HIO engraftment has been well described, here, we aim to elucidate the developmental stages of HIO engraftment and establish if it parallels fetal human intestinal development. A timecourse of transplanted HIOs was analyzed histologically at two, four, six and eight weeks post-transplantation and we demonstrated that the progression of HIO engraftment closely resembles key stages during fetal human intestinal development. We also utilized single nuclear RNA sequencing to determine and track the emergence of distinct cell populations over time and validated our transcriptomic data through in situ protein expression. These rich datasets and observations suggest that transplanted HIOs indeed recapitulate early intestinal development, solidifying their value as a human intestinal model system.

Affiliation: Cincinnati Children's Hosptial Medical Center

Data and Code Availability: Coming soon

References: Seurat

Developed By: Konrad Thorner, Bioinformatics Analyst, Konrad.Thorner@cchmc.org