Analysis of Aged Dysfunctional Intestinal Stem Cells.

Analysis of Aged Dysfunctional Intestinal Stem Cells. Methods Mol Biol. 2020;2171:41-52 Authors: Nalapareddy K, Geiger H Abstract Aging is a multifactorial process. Organ maintenance and tissue regeneration are impaired upon aging mainly due to loss of stem cell function in organs that depend on stem cell in the adult. Intestine is such an organ, and upon aging intestinal regeneration is impaired due to decline of intestinal stem cell function. To determine the aging status of intestine and intestinal stem cells, histological analyses; analyses of the level of proliferation markers in tissue by immunofluorescence and/or quantitative RT-PCR; and gene expression analysis for stemness related genes in isolated crypts, intestinal stem cells (ISC), and Paneth cells can be used. To analyze the level of regeneration in intestine and thus determine a decline in ISC function, techniques like in vitro organoid cultures and lineage tracing with BrdU, lineage tracing using transgenic mice and histological analyses of tissue regeneration after 3 and 5 days after two rounds of 10 Gy of radiation (a 10 + 10 Gy IR experiment) can be applied. In this chapter we will focus on protocols for lineage tracing, the 10 + 10 gy IR experiment and for organoid cultures from young and aged mouse intestine.Lineage tracing experiments in intestine can be done in many ways. In this chapter we describe a protocol for lineage tracing upon BrdU incorporation ...
Source: Mol Biol Cell - Category: Molecular Biology Authors: Tags: Methods Mol Biol Source Type: research