Abstract
Introduction Necrotizing enterocolitis (NEC) is one of the most severe gastrointestinal diseases
in infancy. NEC can cause metabolic derangements, multi-organ injury including severe
liver damage. The mechanism leading to hepatic damage in NEC remains unclear. The
aim of this study is to establish and characterize liver organoids from NEC mice.
Materials and Methods Following ethical approval (#44032), we induced experimental NEC from postnatal day
5 (P5) to P9 using C57BL/6 mice pups. NEC was induced by gavage formula feeding, gavage
lipopolysaccharide (LPS) administration, and hypoxia. Breastfed pups were used as
control. On P9, NEC and control pups were sacrificed and liver tissue was harvested
and organoids were generated. Organoid size was recorded daily (day 2–4) by measuring
their surface area and organoid growth was calculated. RNA was extracted on day 4
after liver organoid generation.
Results Organoid growth rate was significantly lower in NEC liver organoids compared to control
liver organoids. mRNA expression of liver progenitor cells markers of LGR5 and SOX9
was lower in NEC liver organoids compared to control liver organoids. Similarly, expression
of proliferation markers of Ki67 and PCNA was lower in NEC liver organoids.
Conclusion We report a novel technique to generate liver organoids during NEC. These organoids
are characterized by reduced progenitor cells, reduced proliferation, and overall
impaired regenerative capacity. Liver progenitor cells are important targets to prevent
liver damage in NEC and promote recovery.
Keywords
necrotizing enterocolitis - liver injury - organoids - regeneration
