Maternal And Donor Human Milk Support Robust Intestinal Epithelial Growth And Differentiation In A Fetal Intestinal Organoid Model

Author

Lauren SmithFollow

Date of Award

January 2024

Document Type

Open Access Thesis

Degree Name

Medical Doctor (MD)

Department

Medicine

First Advisor

Liza Konnikova

Abstract

Necrotizing enterocolitis (NEC) is a severe gastrointestinal complication of prematurity. Using small intestinal organoids derived from fetal tissue of a gestational age similar to an extremely preterm infant, this study aims to assess the effect of diet on intestinal epithelial growth and differentiation to elucidate the role nutrition type plays in intestinal development and modifies the risk for NEC.

Organoids were cultured for 5 days in organoid growth media and 5 days in organoid differentiation media supplemented 1:40 with four different diet conditions: maternal milk (MM), donor human milk (DHM), standard formula (SF), or extensively hydrolyzed formula (HF). 4x images were captured daily and organoids were quantified via manual counting and measurement. Organoids from both the growth and differentiation phase were preserved for RNA bulk sequencing and immunofluorescence staining with Ki67, chromogranin A, and cleaved caspase 3. Media was saved from day 5 of growth and differentiation for cytokine and growth factor analysis.

Human milk supplementation improved growth and differentiation of fetal intestinal organoids generating larger organoids during the growth phase and organoids with longer and wider buds during differentiation compared to formula.Ki67 staining confirmed the proliferative nature of milk-supplemented organoids and chromogranin A staining proved that MM-supplemented organoids induced highest enteroendocrine differentiation. Human milk supplementation also upregulated genes involved in proliferation and promoted a homeostatic immune landscape while those supplemented with formula had a downregulation of cell- cycle-promoting genes and a more inflammatory immune signature.

Our results show that MM, and to a lesser extent DHM, support robust intestinal epithelial proliferation and differentiation, suggesting a critical role for factors enriched in human milk in intestinal epithelial health.

Comments

This is an Open Access Thesis.

Recommended Citation

Smith, Lauren, "Maternal And Donor Human Milk Support Robust Intestinal Epithelial Growth And Differentiation In A Fetal Intestinal Organoid Model" (2024). Yale Medicine Thesis Digital Library. 4280.
https://elischolar.library.yale.edu/ymtdl/4280