Work

Optimisation of Ex Vivo Peripheral Blood Mononuclear Cell Culture and DNA Double Strand Break Repair Kinetics

Year: 2024

Type: article

Abstract: The assessment and modelling of DNA double-strand break damage and repair is widely investigated throughout the literature. This optimisation study investigated the requirement of cell proliferation prior to treatment with chemotherapeutic agents to damage DNA and the optimal window of analysis for DNA double-strand break repair measurements with γ-H2AX. Peripheral blood mononuclear cells were collected from healthy volunteers and incubated with phytohaemagglutinin at final concentrations of 0, 0.25, 0.5, 1, 2.5, 5 and 10 µg/mL for 0, 24, 48, 72 and 168 h at 37 °C, 5% CO2, and proliferation was measured via spectrometry (MTS assay). This study, detailed in this methodology paper, found that peripheral blood mononuclear cells must be proliferated prior to the chemical induction of DNA double-strand breaks. The window for assessment of early DNA double-strand break repair was determined to be one hour after removal of the DNA damaging agent.

Source: LabMed

DOI: 10.3390/labmed1010003

Authors: Holly Hosking, Wayne Pederick, Paul Neilsen, Andrew Fenning

Institution: Central Queensland University

Language: English

Topic: DNA Repair Mechanisms

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,207