Characterization of conditions and determination of practical tips for mtDNA level estimation in various human cells
Paulina Jędrak , Natalia Sowa , Sylwia Barańska , Grzegorz Węgrzyn
AbstractDetermination 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.
|Journal series||Acta Biochimica Polonica, ISSN 0001-527X, e-ISSN 1734-154X, (A 15 pkt)|
|Publication size in sheets||0.5|
|Keywords in English||mtDNA level, leukocytes, fibroblasts, qPCR methodology, Huntington disease|
|License||Journal (articles only); published final; ; with publication|
|Score|| = 15.0, ArticleFromJournal|
= 15.0, ArticleFromJournal
|Publication indicators||= 2; = 2; : 2017 = 1.239 (2) - 2017=1.541 (5)|
|Citation count*||5 (2019-04-24)|
* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.