Journal de chimie clinique et de médecine de laboratoire

Journal de chimie clinique et de médecine de laboratoire
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Droplet Digital PCR Analysis of miR-191-3p in the Spent Blastocyst Culture Media Might Reflect the Reproductive Competence of the 3rd Day Human Embryo

Katalin Gombos, Miklos Oldal, Kristztina Ildiko Kalacs, Kristztina Godony, Akos Varnagy, Jozsef Bodis and Gabor L Kovacs

Background: Droplet digital PCR analysis of human embryonic culture media collected in a proof of con-cept study on IVF pregnancy outcome. miR-191-3p was analysed in spent human embryonic culture media of morphologically similar, good quality embryos which after later transfer developed either to reproductively competent embryos and healthy neonates or of those, where miscarriage occurred in early stage of the pregnancy. Methods: Spent culture media at the morula-blastocyst stage (3rd day) were collected from embryos fertilised with ICSI and undergoing embryo transfer. After registered pregnancy outcome 40 samples from the group of reproductively competent embryos, 40 samples from miscarriage and their parallel blank culture media samples were enrolled in miRNA analysis. Isolation and quantitative detection of miRNA from embryonic culture media was carried out on automated droplet digital polymerase chain reaction platform. Results: Quantitative analysis and ANOVA evaluation confirmed miR-191-3p to be present in significantly higher concentration in the 3rd day culture media of reproductively competent human embryos (mean concentration difference=20,478, p=1 × 10-4) than the miscarriage. Control blank culturing media were negative for miR-191-3p. Conclusion: miR-191-3p present in human blastocyst culture media indicates actual embryonic origin and changed in expression patterns depending on clinical outcome after transfer. It might be a candidate molecular marker for pre-implantation assessment of reproductive embryo competence in a non-invasive way.