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@ARTICLE{Michel:134835,
author = {C. Michel$^*$ and R. Küchler$^*$ and I. Martin$^*$ and B.
Kyewski$^*$ and S. Pinto$^*$},
title = {{A}n efficient protocol for in vivo labeling of
proliferating epithelial cells.},
journal = {Journal of immunological methods},
volume = {457},
issn = {0022-1759},
address = {Amsterdam [u.a.]},
publisher = {Elsevier Science},
reportid = {DKFZ-2018-00625},
pages = {82 - 86},
year = {2018},
abstract = {The study of organogenesis, tissue-homeostasis and
regeneration requires the precise assessment of in vivo cell
proliferation. To this end a host of methods have been
developed to detect and quantify DNA synthesis in
proliferating cells. These include cell labeling with
various nucleotide analogues and fluorescence reporter-based
animal models with each method presenting its idiosyncratic
shortcomings. Quantitative assessment of epithelial cell
turnover has been partly hampered due to their variable and
limited in vivo accessibility and the requirement for
harsher isolation procedures to procure single cells for
FACS analysis. Here, we report a reliable protocol to study
in vivo cell proliferation of epithelial cells in mice by
repeatedly injecting EdU intravenously for an extended
12-day period. EdU incorporation was quantitated ex vivo by
FACS after tissue dissociation in order to obtain single
epithelial cell suspensions. As a lead population, we
analyzed thymic epithelial cells (TECs), where we were able
to label compartmentalized TEC subsets to saturation without
apparent toxic effects on the thymus architecture or
stress-sensitive TEC lineage differentiation. The data is in
concordance with the prevailing model of medullary TEC
terminal differentiation that includes the post-Aire stage.
The same protocol was successfully applied to epithelial
cells of various other organs - skin, lymph node, kidney and
small intestine - tissues with widely varying frequencies
and rates of proliferating epithelial cells.},
cin = {D090 / A110},
ddc = {610},
cid = {I:(DE-He78)D090-20160331 / I:(DE-He78)A110-20160331},
pnm = {314 - Tumor immunology (POF3-314)},
pid = {G:(DE-HGF)POF3-314},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:29605230},
doi = {10.1016/j.jim.2018.03.015},
url = {https://inrepo02.dkfz.de/record/134835},
}