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Related Concept Videos

Renewal of Intestinal Stem Cells01:23

Renewal of Intestinal Stem Cells

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The intestinal epithelial lining rapidly renews every 4 to 5 days. The renewal is facilitated by intestinal stem cells (ISCs) located at the base of the crypt– a gland located at the bottom of each villus. ISCs divide asymmetrically to form new stem cells and progenitor daughter cells. The daughter cells are called transit-amplifying (TA) cells which move upwards along the crypt and either differentiate into absorptive cells– the enterocytes or secretory cells– including the...
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Adult Stem Cells01:33

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Stem cells are undifferentiated cells that divide and produce more stem cells or progenitor cells that differentiate into mature, specialized cell types. All the cells in the body are generated from stem cells in the early embryo, but small populations of stem cells are also present in many adult tissues including the bone marrow, brain, skin, and gut. These adult stem cells typically produce the various cell types found in that tissue—to replace cells that are damaged or to continuously...
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Role Of Notch Signalling In Intestinal Stem Cell Renewal01:12

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Notch signaling was first discovered in Drosophila melanogaster, where it is involved in cell lineage differentiation. Notch signaling regulates the maintenance and differentiation of intestinal stem cells or ISCs by controlling the expression of atonal homolog 1 or Atoh1. Atoh1 directs cells to differentiate into secretory cells.
Direct cell-to-cell contact is needed for the activation of Notch signaling. The signal is initiated when a notch ligand binds to a receptor on an adjacent cell, also...
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Stem Cell Culture01:17

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Stem cell research aims to find ways to use stem cells to regenerate and repair cellular damage. Over time, most adult cells undergo the wear and tear of aging and lose their ability to divide and repair themselves. Stem cells do not display a particular morphology or function. Adult stem cells, which exist as a small subset of cells in most tissues, keep dividing and can differentiate into a number of specialized cells generally formed by that tissue. These cells enable the body to renew and...
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Related Experiment Video

Updated: Dec 14, 2025

Isolating Intestinal Stem Cells from Adult Drosophila Midguts by FACS to Study Stem Cell Behavior During Aging
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Analysis of Aged Dysfunctional Intestinal Stem Cells.

Kodandaramireddy Nalapareddy1, Hartmut Geiger2

  • 1Division of Experimental Hematology, Cancer Biology & Stem Cell Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA.

Methods in Molecular Biology (Clifton, N.J.)
|July 25, 2020
PubMed
Summary

Aging impairs intestinal regeneration due to declining intestinal stem cell (ISC) function. This study details methods like lineage tracing and organoid cultures to quantify this age-related decline in ISC regenerative capacity.

Keywords:
AgingIntestinal stem cellsIrradiationLgr5MiceOrganoidsRegeneration

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The Organoid Reconstitution Assay ORA for the Functional Analysis of Intestinal Stem and Niche Cells
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The Organoid Reconstitution Assay ORA for the Functional Analysis of Intestinal Stem and Niche Cells

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Area of Science:

  • Gastroenterology
  • Stem Cell Biology
  • Gerontology

Background:

  • Organ maintenance and tissue regeneration decline with age, primarily due to diminished stem cell function.
  • Intestinal regeneration is particularly affected by aging, linked to a decline in intestinal stem cell (ISC) function.
  • Quantifying age-related changes in ISC function is crucial for understanding intestinal aging.

Purpose of the Study:

  • To present protocols for assessing intestinal stem cell function in aged versus young mice.
  • To detail methods for evaluating intestinal regeneration and ISC function decline.
  • To provide a framework for analyzing the impact of aging on intestinal stem cells.

Main Methods:

  • Lineage tracing using Bromodeoxyuridine (BrdU) incorporation and Lgr5-EGFP-CreERT2; Rosa26-YFP transgenic mice.
  • In vivo regeneration assessment via a 10 Gy + 10 Gy radiation (10+10 Gy IR) experiment analyzing crypt depth and fission.
  • In vitro intestinal organoid cultures from young and aged mouse crypts, including serial replating to assess stem cell function.

Main Results:

  • Reduced frequency of organoid formation and fewer lobes per organoid were observed in aged mice after serial replating.
  • The described lineage tracing and radiation models allow for quantitative assessment of ISC regenerative potential.
  • Histological analyses of crypt depth and fission rates provide in vivo estimates of regenerative capacity.

Conclusions:

  • The combination of lineage tracing, radiation-induced regeneration assays, and organoid cultures effectively quantifies age-related decline in ISC function.
  • These methods demonstrate a significant reduction in the regenerative potential of intestinal stem cells with aging.
  • The presented protocols offer a robust approach to studying intestinal stem cell aging and regeneration.