Isolation of senescent cells by iodixanol (OptiPrep) density gradient-based separation

Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00159816%3A_____%2F19%3A00072490" target="_blank" >RIV/00159816:_____/19:00072490 - isvavai.cz</a>
Result on the web
<a href="https://onlinelibrary.wiley.com/doi/pdf/10.1111/cpr.12674" target="_blank" >https://onlinelibrary.wiley.com/doi/pdf/10.1111/cpr.12674</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1111/cpr.12674" target="_blank" >10.1111/cpr.12674</a>

Result language
angličtina
Original language name
Isolation of senescent cells by iodixanol (OptiPrep) density gradient-based separation
Original language description
Objectives Chemotherapeutic drugs induce senescence in cancer cells but, unlike replicative senescence or oncogene-induced senescence, do so rather inefficiently and depending on DNA damage. A thorough understanding of the biology of chemotherapy-induced senescent cells requires their isolation from a mixed population of adjacent senescent and non-senescent cancer cells. Materials and methods We have developed and optimized a rapid iodixanol (OptiPrep)-based gradient centrifugation system to identify, isolate and characterize doxorubicin (DXR)-induced senescent hepatocellular carcinoma (HCC) cells (HepG2 and Huh-7) in vitro. Results After cellular exposure to DXR, we used iodixanol gradient-based centrifugation to isolate and re-plate cells on collagen-coated flasks, despite their low or null proliferative capacity. The isolated cell populations were enriched for DXR-induced senescent HCC cells, as confirmed by proliferation arrest assay, and beta-galactosidase and DNA damage-dependent gamma H2A.X staining. Conclusions Analysing pure cultures of chemotherapy-induced senescent versus non-responsive cancer cells will increase our knowledge on chemotherapeutic mechanisms of action, and help refine current therapeutic strategies.
Czech name
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Czech description
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Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10601 - Cell biology

Project
<a href="/en/project/EF15_003%2F0000492" target="_blank" >EF15_003/0000492: Unveiling the molecular determinants of agingto design new therapeutics</a><br>
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year
2019
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

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