Characterization of conditions and determination of practical tips for mtDNA level estimation in various human cells

• Authors: Paulina Jędrak , Natalia Sowa , Sylwia Barańska , Grzegorz Węgrzyn
• Languages of publication: EN

Abstracts

EN

Determination of mtDNA copy number in the cell is crucial to understand many cellular processes. Recently, the number of studies with the use of mitochondrial DNA (mtDNA) content as the determinant of mitochondrial abnormalities increased greatly and is still growing, therefore, optimization of technical conditions for this analysis is crucial. Despite using similar laboratory protocols, some results cannot be compared between research centers, thus causing discrepancies in the assessment of mtDNA content. The aim of this work was to test which conditions of biological sample collection and storage affect estimation of mtDNA level relative to the nuclear DNA (nDNA) in the blood samples and dermal fibroblasts. We found that the time and temperature of sample storage, as well as the type of the blood sample (whole blood or leukocytes) influence the estimate of mtDNA/nDNA ratio in the blood. In the case of dermal fibroblasts collected from healthy control and Huntington disease patients, our data indicate that the passage number of cells is essential to obtain reliable results.

Keywords

EN

mtDNA level; leukocytes; fibroblasts; qPCR methodology; Huntington disease

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2017
• Volume: 64
• Issue: 4
• Pages: 699-704

Dates

published

2017

received

2017-08-17

revised

2017-11-13

accepted

2017-11-14

(unknown)

2017-12-15

Contributors

author

Paulina Jędrak

• Department of Molecular Biology, University of Gdańsk, Gdańsk, Poland

author

Natalia Sowa

• Department of Molecular Biology, University of Gdańsk, Gdańsk, Poland

author

Sylwia Barańska

• Department of Molecular Biology, University of Gdańsk, Gdańsk, Poland

author

Grzegorz Węgrzyn

• Department of Molecular Biology, University of Gdańsk, Gdańsk, Poland

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Identifiers

DOI

10.18388/abp.2017_2303