Aging effects on intestinal homeostasis associated with expansion and dysfunction of intestinal epithelial stem cells

Aging (Albany NY). 2017 Aug 29;9(8):1898-1915. doi: 10.18632/aging.101279.


Intestinal epithelial stem cells (IESCs) are critical to maintain intestinal epithelial function and homeostasis. We tested the hypothesis that aging promotes IESC dysfunction using old (18-22 months) and young (2-4 month) Sox9-EGFP IESC reporter mice. Different levels of Sox9-EGFP permit analyses of active IESC (Sox9-EGFPLow), activatable reserve IESC and enteroendocrine cells (Sox9-EGFPHigh), Sox9-EGFPSublow progenitors, and Sox9-EGFPNegative differentiated lineages. Crypt-villus morphology, cellular composition and apoptosis were measured by histology. IESC function was assessed by crypt culture, and proliferation by flow cytometry and histology. Main findings were confirmed in Lgr5-EGFP and Lgr5-LacZ mice. Aging-associated gene expression changes were analyzed by Fluidigm mRNA profiling. Crypts culture from old mice yielded fewer and less complex enteroids. Histology revealed increased villus height and Paneth cells per crypt in old mice. Old mice showed increased numbers and hyperproliferation of Sox9-EGFPLow IESC and Sox9-EGFPHigh cells. Cleaved caspase-3 staining demonstrated increased apoptotic cells in crypts and villi of old mice. Gene expression profiling revealed aging-associated changes in mRNAs associated with cell cycle, oxidative stress and apoptosis specifically in IESC. These findings provide new, direct evidence for aging associated IESC dysfunction, and define potential biomarkers and targets for translational studies to assess and maintain IESC function during aging.

Keywords: Sox9; aging; intestinal epithelial stem cells; organoid.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Age Factors
  • Aging / genetics
  • Aging / metabolism
  • Aging / pathology*
  • Animals
  • Apoptosis
  • Cell Cycle
  • Cell Lineage
  • Cell Proliferation*
  • Enterocytes / metabolism
  • Enterocytes / pathology
  • Epithelial Cells / metabolism
  • Epithelial Cells / pathology*
  • Female
  • Gene Expression Regulation, Developmental
  • Genotype
  • Goblet Cells / metabolism
  • Goblet Cells / pathology
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Homeostasis
  • Intestinal Mucosa / metabolism
  • Intestinal Mucosa / pathology*
  • Jejunum / metabolism
  • Jejunum / pathology*
  • Lac Operon
  • Male
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Oxidative Stress
  • Paneth Cells / metabolism
  • Paneth Cells / pathology
  • Phenotype
  • Receptors, G-Protein-Coupled / genetics
  • Receptors, G-Protein-Coupled / metabolism
  • SOX9 Transcription Factor / genetics
  • SOX9 Transcription Factor / metabolism
  • Signal Transduction
  • Spheroids, Cellular
  • Stem Cells / metabolism
  • Stem Cells / pathology*
  • Time Factors
  • Tissue Culture Techniques


  • Lgr5 protein, mouse
  • Receptors, G-Protein-Coupled
  • SOX9 Transcription Factor
  • Sox9 protein, mouse
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins