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Simple, fast, cost-efficient, reliable, and highly automated DNA content analysis of cells in adherent cultures

J. Čížková, A. Filipová, A. Carrillo, M. Ehrlichová, A. Spálenková, A. Magdolenová, M. Hájek, P. Horák, A. Erbenova, Z. Šinkorová

. 2024 ; 105 (6) : 474-479. [pub] 20240503

Jazyk angličtina Země Spojené státy americké

Typ dokumentu časopisecké články

Perzistentní odkaz   https://www.medvik.cz/link/bmc24013652

Grantová podpora
Ministerstvo Obrany České Republiky
Ministerstvo Zdravotnictví Ceské Republiky
Ministerstvo Školství, Mládeže a Tělovýchovy
Grantová Agentura České Republiky

E-zdroje Online Plný text

NLK Free Medical Journals od 2003 do Před 1 rokem
Medline Complete (EBSCOhost) od 2012-06-01 do Před 1 rokem
Wiley Free Content od 2003 do Před 1 rokem

Odkazy

PubMed 38702936
DOI 10.1002/cyto.a.24840
Knihovny.cz E-zdroje

The most commonly used flow cytometric (FCM) analysis of cellular DNA content relies on ethanol fixation followed by RNA digestion and propidium iodide (PI) intercalation into double-stranded DNA. This is a laborious and time-consuming procedure that is subject to systematic errors due to centrifugation and washing steps associated with sample preparation. It can adversely affect the reliability of the results. Here, we present a modified concept of DNA quantification in adherent cell lines by FCM that involves neither ethanol fixation nor any washing and cell transferring steps. Our high throughput assay of adherent cell lines reduces sample-processing time, requires minimal workload, provides a possibility for automation, and, if needed, also allows a significant reduction in the size of individual samples. Working with a well-proven commercial tool-The BD CycletestTM Plus DNA Reagent Kit-primarily designed for cell cycle analysis and aneuploidy determination in experimental and clinical samples, we suggest a novel, very efficient, and robust approach for DNA research in adherent cell cultures.

Citace poskytuje Crossref.org

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