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The remaining authors have no conflicts of interest to declare. receives support from the NIHR Oxford Biomedical Research Centre. holds US20170039415A1, both for metabolic imaging methods. This study was funded by National Institutes of Health grant 5R01HD092550-02. Nonetheless, our data raise interesting biological questions, particularly regarding the interplay of metabolism and BMI and the association of mtDNA haplogroup with oocyte yield in ovarian stimulation cycles. These findings suggest that the analysis of mtDNA in CCs is unlikely to provide an advantage in terms of improved embryo selection during assisted reproduction cycles. Also, the results obtained with respect to specific clinical outcomes and macrohaplogroups should be interpreted with caution due to the smaller sample sizes when subdividing the dataset. There was a non-significant increase in mtDNA levels in CCs with heteroplasmic mitochondrial mutations.Īlthough a large number of samples were analysed in this study, it was not possible to analyse all the CCs from every patient.
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The total number of retrieved oocytes was different between the H-V and J-T as well as the U-K and J-T macrohaplogroups. The number of variants predicted to affect the ATP production differed significantly between mitochondrial macrohaplogroups. The COXI locus contained the least number of coding sequence variants, whereas ATPase8 had the most. mtGenome sequencing revealed 4181 genetic variants with respect to a reference genome. No significant associations were observed between CC mtDNA quantity and patient age, female infertility diagnosis or any ART outcome variable. The relative mtDNA CC content was significantly associated with BMI. Sequence variants in the mtDNA were evaluated using Mitomaster and HmtVar to predict their potential impact. Massively parallel sequencing permitted not only the accurate detection of mutations but also the precise quantification of levels of mutations in cases of heteroplasmy.
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Associations between relative mtDNA quantity and mtGenome variants in CCs and patient age, BMI (kg/m2), infertility diagnosis and ART outcomes were investigated. mtDNA quantity was measured in all the samples with a validated quantitative PCR method and the entire mtGenome was sequenced in a subset of 138 samples using a high-depth massively parallel sequencing approach. This was a prospective observational study involving human CCs collected with 541 oocytes from 177 IVF patients. Currently, little is known concerning variation in CC mitochondrial genetics and how this might influence the reproductive potential of the associated oocyte. Such mutations may be homoplasmic (affecting all mtDNA in a cell) or they may display varying levels of heteroplasmy (affecting a proportion of the mtDNA). mtDNA mutations can be inherited or they can accumulate in somatic cells over time, potentially contributing to aging. As with other cell types, appropriate cellular function is likely to rely upon adequate energy production, which in turn depends on the quantity and genetic competence of the mitochondria. Neither the CC mtDNA quantity nor the presence of specific mtDNA genetic variants was associated with ART outcomes, although associations with patient body mass index (BMI) were detected, and the total number of oocytes retrieved differed between major mitochondrial haplogroups.ĬCs fulfil a vital role in the support of oocyte developmental competence. Are relative mitochondrial DNA (mtDNA) content and mitochondrial genome (mtGenome) variants in human cumulus cells (CCs) associated with oocyte reproductive potential and assisted reproductive technology (ART) outcomes?
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