Optimization of microRNA Acquirement from Seminal Plasma and Identification of Diminished Seminal microRNA-34b as Indicator of Low Semen Concentration.

International Journal of Molecular Sciences
Michael EikmansFrans H J Claas

Abstract

About 10-15% of couples who want to conceive suffer from subfertility, while in 30% of these cases, a male factor plays a role. Levels of particular microRNAs in seminal plasma, including those involved in spermatogenesis, may serve as an indicative parameter for subfertility. We first optimized a protocol for acquiring microRNAs from seminal plasma. Next, using a test-validation strategy in a male cohort, we aimed to identify microRNAs of which the levels are related to semen motility and concentration. By qPCR, 742 microRNAs were profiled in three normozoospermic samples, three seminal samples with a low semen motility (asthenozoospermia), and three with a low semen concentration (oligozoospermia). MicroRNAs showing significant differences between groups were further validated in a second cohort consisting of 40 samples with normozoospermia (control group), 47 samples with asthenozoospermia, and 19 samples with oligozoospermia (of which 74% also low motility). Highest microRNA yields were obtained with the Biofluids RNA extraction kit, with inclusion of MS2 RNA carrier and proteinase K treatment to the protocol, and when 50 µL of seminal plasma was used as input. Exosome isolation prior to RNA extraction did not lead to enhan...Continue Reading

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Citations

Nov 18, 2020·International Journal of Molecular Sciences·Fengli WangShuiqiao Yuan
Apr 4, 2021·International Journal of Molecular Sciences·Madalina Gabriela BarbuDragos Valentin Predescu
Apr 4, 2021·International Journal of Molecular Sciences·Manuel Álvarez-Rodríguez, Felipe Martinez-Pastor
Nov 6, 2021·Andrologia·Reza Asadpour, Ehsan Mofidi Chelan

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Methods Mentioned

BETA
PCR
biopsies

Software Mentioned

Exiqon

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