A novel human epithelial enteroid model of necrotizing enterocolitis

Guillermo J. Ares, Christie Buonpane, Carrie Yuan, Douglas Wood, Catherine J. Hunter*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

20 Scopus citations


Necrotizing enterocolitis (NEC) is a devastating disease of newborn infants. It is characterized by multiple pathophysiologic alterations in the human intestinal epithelium, leading to increased intestinal permeability, impaired restitution, and increased cell death. Although there are numerous animal models of NEC, response to injury and therapeutic interventions may be highly variable between species. Furthermore, it is ethically challenging to study disease pathophysiology or novel therapeutic agents directly in human subjects, especially children. Therefore, it is highly desirable to develop a novel model of NEC using human tissue. Enteroids are 3-dimensional organoids derived from intestinal epithelial cells. They are ideal for the study of complex physiologic interactions, cell signaling, and host-pathogen defense. In this manuscript we describe a protocol that cultures human enteroids after isolating intestinal stem cells from patients undergoing bowel resection. The crypt cells are cultured in media containing growth factors that encourage differentiation into the various cell types native of the human intestinal epithelium. These cells are grown in a synthetic, collagenous mix of proteins that serve as a scaffold, mimicking the extra-cellular basement membrane. As a result, enteroids develop apical-basolateral polarity. Co-administration of lipopolysaccharide (LPS) in media causes an inflammatory response in the enteroids, leading to histologic, genetic, and protein expression alterations similar to those seen in human NEC. An experimental model of NEC using human tissue may provide a more accurate platform for drug and treatment testing prior to human trials, as we strive to identify a cure for this disease.

Original languageEnglish (US)
Article numbere59194
JournalJournal of Visualized Experiments
Issue number146
StatePublished - Apr 2019


  • Enteroid
  • Epithelial organoid
  • Human tissue model
  • Immunology and Infection
  • Intestinal crypt
  • Issue 146
  • LPS model
  • Necrotizing Enterocolitis
  • Stem cells

ASJC Scopus subject areas

  • General Chemical Engineering
  • General Immunology and Microbiology
  • General Biochemistry, Genetics and Molecular Biology
  • General Neuroscience


Dive into the research topics of 'A novel human epithelial enteroid model of necrotizing enterocolitis'. Together they form a unique fingerprint.

Cite this