# Enteroid Core

> **NIH NIH P01** · UNIVERSITY OF MARYLAND BALTIMORE · 2024 · $396,399

## Abstract

ENTEROID CORE – Project Summary
 The Enteroid Core (EC) is a multi-service resource that provides Program Project and Core Investigators and
their laboratories the tools and advice needed to establish and utilize human intestinal stem cell-derived
enteroids/colonoids for understanding the pathogenesis of enteric bacterial infections. The Enteroid Core
provides established enteroid and colonoid cultures from JHU biobank, expertise and training in the use of human
small intestinal and colonic enteroid monolayers, growth factor-conditioned media required for propagation,
maintenance, and differentiation of enteroid cultures, including M cells, as well as assistance in troubleshooting
growth, differentiation, and use of enteroids for study of host-pathogen interactions in each project. The Enteroid
Core works in close collaboration with the Immunology Core to develop novel co-culture models of human
enteroids/colonoids with primary human innate immune cells. Our collaborative work during the current award
period resulted in the first human immune-enteroid co-culture model that included monocyte-derived
macrophages. In addition, the Enteroid Core developed techniques and protocols to assist Project laboratories
achieve their proposed Aims including lentiviral transduction of human enteroids/colonoids for stable knockdown
(via shRNA) of target genes (e.g. MUC2 for Project 1 (EAEC); MRP5 for Project 3 (ETEC)) and differentiation of
specialized intestinal epithelial cell types (e.g. M cells for Project 2 (Shigella)). This renewal application seeks to
continue these collaborative efforts to provide Projects 2 (Shigella) and 3 (ETEC) and the Immunology Core with
media necessary for propagating human enteroids/colonoids and growing them as monolayers as well as media
to normally differentiate enteroids/colonoids as well as to express M cells (Project 2 (Shigella)). Project 1 (EAEC)
investigators have been trained to make conditioned media independently and will receive quality control support
from the EC. The EC will continue to help with troubleshooting problems with culturing enteroids as well as use
of enteroids for each project. In collaboration with the Immunology Core, the EC will continue developing innate
immune cell-enteroid co-culture models to include human monocyte-derived dendritic cells and intraepithelial
lymphocytes. In addition to new co-culture models, the EC and Immunology Core will increase the complexity of
co-cultures through increased direct physical interaction of enteroid monolayers with immune cells normally
present in the lamina propria, using commercially available 3D cell culture scaffolds, perfusion systems, and
recently developed anaerobic chamber used for human enteroid monolayers.

## Key facts

- **NIH application ID:** 10868675
- **Project number:** 5P01AI125181-09
- **Recipient organization:** UNIVERSITY OF MARYLAND BALTIMORE
- **Principal Investigator:** Nicholas Constantine Zachos
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $396,399
- **Award type:** 5
- **Project period:** 2016-07-01 → 2026-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10868675

## Citation

> US National Institutes of Health, RePORTER application 10868675, Enteroid Core (5P01AI125181-09). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10868675. Licensed CC0.

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