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

Kód výsledku v IS VaVaI

RIV/00159816:_____/19:00072490 - isvavai.cz

Výsledek na webu

https://onlinelibrary.wiley.com/doi/pdf/10.1111/cpr.12674

DOI - Digital Object Identifier

10.1111/cpr.12674

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10601 - Cell biology

Projekt

EF15_003/0000492: Mapování molekulární podstaty procesů stárnutí pro vývoj nových léčebných metod

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2019

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

CELL PROLIFERATION

ISSN

0960-7722

e-ISSN

—

Svazek periodika

52

Číslo periodika v rámci svazku

6

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

9

Strana od-do

—

Kód UT WoS článku

000486414300001

EID výsledku v databázi Scopus

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