Induction of Necrotizing Enterocolitis in Human Epithelial Enteroid Model

Abstract

Source: Ares, G. J., et al. A Novel Human Epithelial Enteroid Model of Necrotizing Enterocolitis. J. Vis. Exp.(2019).

This video demonstrates the development of necrotizing enterocolitis disease in a human-derived neonatal enteroid model induced by lipopolysaccharides treatment. This model helps to study intestinal pathophysiology.

Protocol

All procedures involving human participants have been performed in compliance with the institutional, national, and international guidelines for human welfare and have been reviewed by the local institutional review board.

1. Reagent Preparation

1. Prepare Culture Media stock solution for whole tissue collection: 1 L of Dulbecco's Modified Eagle Medium (DMEM), 110 mL of Fetal Bovine Serum (FBS), 11 mL of Penicillin-Streptomycin (final concentration 1%), and 1.1 mL of filter-sterilized insulin (final concentration 0.1%.) Store stock solution at 4 °C.
2. Prepare Chelating Buffer #1: 30 mL of Culture Media....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
4% Paraformaldehyde	ThermoFisher	AAJ19943K2
Basement Membrane Matrix (Matrigel)	Corning	CB-40230C
DMEM/F-12	ThermoFisher	MT-16-405-CV
Dulbecco's Modified Eagle Medium (DMEM)	ThermoFisher	11-965-118
Dulbecco's Phosphate-Buffered Saline (DPBS)	ThermoFisher	14190-144
Epidermal Growth Factor (EGF)	Sigma	E9644-.2MG
Ethylenediaminetetraacetic acid (EDTA)	Sigma	EDS-500G
Fetal Bovine Serum (FBS)	Gemini Bio-Pro	100-125
Phosphate Buffered Saline (PBS)	Sigma	P5368-5X10PAK
RPMI 1640 Medium	Invitrogen	11875093
Tissue Processing Gel (Histogel)	ThermoFisher	22-110-678
Lipopolysaccharide (LPS)	Sigma	L2630-25MG
Gentamicin	Sigma	G5013-1G