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

Paulina Jędrak , Natalia Sowa , Sylwia Barańska , Grzegorz Węgrzyn

Abstract

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.
Author Paulina Jędrak KBM
Paulina Jędrak ,,
- Department of Molecular Biology
, Natalia Sowa WB
Natalia Sowa,,
- Faculty of Biology
, Sylwia Barańska KBM
Sylwia Barańska,,
- Department of Molecular Biology
, Grzegorz Węgrzyn KBM
Grzegorz Węgrzyn,,
- Department of Molecular Biology
Journal seriesActa Biochimica Polonica, ISSN 0001-527X, e-ISSN 1734-154X
Issue year2017
Vol64
No4
Pages699-704
Publication size in sheets0.5
Keywords in EnglishmtDNA level, leukocytes, fibroblasts, qPCR methodology, Huntington disease
DOIDOI:10.18388/abp.2017_2303
URL https://doi.org/10.18388/abp.2017_2303
Languageen angielski
Score (nominal)15
ScoreMinisterial score = 15.0, 26-02-2018, ArticleFromJournal
Ministerial score (2013-2016) = 15.0, 26-02-2018, ArticleFromJournal
Publication indicators WoS Impact Factor: 2016 = 1.159 (2) - 2016=1.491 (5)
Citation count*0
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* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.
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