Abstract
DNA damage in cumulus cells (CCs) might be related with the developmental competence of the enclosed oocytes, however, conclusivestudies are missing, partially due to the lack of a reliable, cheap, fast, and reproducible DNA damage test. We report the development of a chromatin dispersion test that allows for a fast evaluation of double strand DNA (ds-DNA) damage in CCs. The whole experiment was performed using CCs from 103 oocyte retrieval cycles evaluating the prototype D3-MAX ability (a chromatin dispersion based assay) to detect DNA breaks against in situ nick translation (ISNT) and a two tailed comet assay (TT-comet). Samples were collected from womenyounger than 35 years of age with a good response to stimulation. Pooled cumulus cells of MII oocytes were used. The chromatin dispersion assay results correlate with the double strand-DNA breaks values assessed by the TT-comet assay (Spearman Rho = 0.624; p = 0.003;), while the correlation was poor when compared to the single strand DNA (ss-DNA) breaks observed also with the TT-comet assay (Spearman Rho = -0.141; p = 0.554). ISNT showed a correspondence in the same cells between enzymatic incorporation of modified nucleotides andhalos of chromatin dispersion. We conclude that D3-Max test detects mainly ds-DNA breaks in cumulus cells and is a reliable, fast, and easy reproducible assay suitable for routine clinical practices once the influence on oocyte quality has been established.
KEYWORDS:
Assisted reproduction; DNA fragmentation; chromatin dispersion test; cumulus cells
Year: 2015
Journal: Systems Biology in Reproductive Medicine
PMID: 26305732
Tags: assisted reproduction, chromatin dispersion test, cumulus cells, dna fragmentation
Centers: Congratulations to Aina Nicolau for defending her Master’s Thesis New publication in Molecular Human Reproduction New publication in the Journal of Assisted Reproduction and Genetics Agustina Alessandroni has joined our laboratory to pursue her PhD degree.