Granulocyte-Macrophage Colony Stimulating Factor in Single Blastocyst Conditioned Medium as a Biomarker for Predicting Implantation Outcome of Embryo

Abstract

BackgroundIt is highly desirable to develop new strategies based on secretomics to more accurately selection of embryos with the highest developmental potential for transfer. Granulocyte-macrophage colony-stimulating factor (GM-CSF) has been reported to promote embryo development and pregnancy establishment. However, the predictive value of GM-CSF in single blastocyst selection remains unclear. This study is to determine the concentration of GM-CSF in human single-blastocyst conditioned medium (SBCM) and to evaluate its association with embryo quality and pregnancy outcome.MethodsThe patients with ≤38 years of age receiving the first cycle of assisted reproductive therapy were included in this study. The patients who had <4 top-quality embryos formed by the fertilized two pronuclear zygotes on day 3 were excluded. A total of 126 SBCM samples (SBCMs) were included, of which blastocysts from 77 SBCMs were later transferred in subsequent frozen-thawed embryo transfer. The concentrations of GM-CSF were detected by single-molecule array (SIMOA) and analyzed for their possible association with embryo quality and pregnancy outcomes. The top-quality embryo (TQ), positive HCG (HP), clinical pregnancy (CP), and ongoing pregnancy (OP) rates were determined and compared between groups divided based on GM-CSF concentrations.ResultsThe detection rate of GM-CSF was found to be 50% in all SBCMs. There were significant differences in TQ rate, HP rate, CP rate and OP rate among high concentration group, medium concentration group and low concentration group. Both GM-CSF alone or GM-CSF combined with the morphological score (MS) had a greater AUC of ROC curve than that of MS alone to predict the pregnancy outcome, and GM-CSF combined with MS had the highest AUC.ConclusionsThe concentration of GM-CSF in SBCM was detected at fg/ml levels, which was associated with embryo quality and pregnancy outcome. Collectively, GM-CSF may be used as a biomarker for prediction of pregnancy outcome and selection of embryos with high developmental potential for transfer in assisted reproductive technology (ART).

Molecular regulators of human blastocyst development and hatching: Their significance in implantation and pregnancy outcome

“…41,43,102 Potential embryo-viability biomarkers in embryo secretome or embryo spent medium (E-SM), associated with improved IVF outcomes, reported so far include, sHLA-G, 49,50,83,86 microRNAs, 103 telomeric repeat-containing RNAs (TERRRA), 104 ubiquitin, 51,52,100 apolipoprotein-A, 105 platelet-activating factor (PAF), 106 human haptoglobin molecule, 107 human chorionic gonadotrophin (hCG), 108 mitochondria, 109 and mitochondrial DNA content, 110 Glutamine 53 and cytokines such as IL-1β, 72 tumour necrosis factor α (TNF-α), 72,111 LIF 111,112 and Granulocyte-macrophage colony stimulating factor (GM-CSF). 113 Among these identified embryo-derived biomolecules, sHLA-G and ubiquitin could be used as potential predictive biomarkers for improved LBR. They show a strong positive correlation with embryo developmental stage, their quality scores and LBR.…”

“…51,52 Though, earlier studies on embryo biomarkers have showed their association with the embryo quality and pregnancy outcome. 49,50,53,72,83,86,89,113 they are still restricted on embryo viability biomarker estimations in E-SMs of morulae (day 4) and blastocyst stage (days 5-6), and their correlative analysis with pregnancy outcome in terms of LBR, following their ETs. Also, most studies so far have measured other biomarkers in only cleavage-stage embryos.…”

“…Also, most studies so far have measured other biomarkers in only cleavage-stage embryos. 49,53,83,86,89,113 Besides, in global context and also in India, to the best of our knowledge, there has not been a single study on biomarker estimations in human E-SMs from SETs and report as predictive biomarker for pregnancy outcome, despite a huge number of IVF-ET cycles performed. This is despite the fact that a significant research has been carried out on the retrospective correlative studies on embryo-viability biomarkers and pregnancy outcome.…”

“…they are still restricted on embryo viability biomarker estimations in E‐SMs of morulae (day 4) and blastocyst stage (days 5–6), and their correlative analysis with pregnancy outcome in terms of LBR, following their ETs. Also, most studies so far have measured other biomarkers in only cleavage‐stage embryos 49,53,83,86,89,113 . Besides, in global context and also in India, to the best of our knowledge, there has not been a single study on biomarker estimations in human E‐SMs from SETs and report as predictive biomarker for pregnancy outcome, despite a huge number of IVF‐ET cycles performed.…”

Cathepsin-L Secreted by High-Quality Bovine Embryos Exerts an Embryotrophic Effect In Vitro

“…Several studies focused on the secretion of immunological factors by human and murine embryos, as they play a major role in the embryo-maternal dialogue by facilitating immune-related implantation events (for review, see Bahrami-Asl [ 25 ]). Granulocyte–macrophage colony-stimulating factor, a pertinent regulator for the immune response during implantation, was detected in the secretome of both human [ 36 ] and murine [ 37 ] embryos, and was considered a potential marker of viable blastocysts [ 36 , 37 , 38 ]. Katz-Jaffe and colleagues [ 31 ] reported similar protein profiles between the human and murine secretome, and proposed the regulatory protein ubiquitin as a candidate biomarker for blastocyst development in both species [ 31 ].…”