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Up-regulated microRNAs in blastocoel fluid of human implanted embryos could control circuits of pluripotency and be related to embryo competence

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JOURNAL OF ASSISTED REPRODUCTION AND GENETICS 2025年 1-15页

作者： Battaglia, Rosalia Caponnetto, Angela Ferrara, Carmen Fazzio, Anna Barbagallo, Cristina Stella, Michele Barbagallo, Davide Ragusa, Marco Vento, Maria Elena Borzi, Placido Scollo, Paolo Carli, Luca Feichtinger, Michael Kasapi, Evangelia Tsakos, Elias Palini, Simone Sierka, Wojciech Pecorino, Basilio Campitiello, Maria Rosaria Ronsini, Carlo Purrello, Michele Valerio, Domenico Longobardi, Salvatore D'Hooghe, Thomas Di Pietro, Cinzia Univ Catania Dept Biomed & Biotechnol Sci Sect Biol & Genet G Sichel I-95123 Catania Italy Univ Enna Kore Dept Med & Surg I-94100 Enna Italy Cannizzaro Hosp IVF Unit Catania Italy Wunschbaby Inst Feichtinger Vienna Austria EmbryoClin IVF Thessaloniki Greece Cervesi Hosp IVF Unit Cattolica Italy Gyncentrum Sp Zelazna 1 PL-40851 Katowice Poland ASL Salerno Dept Obstet & Gynecol & Physiopathol Human Reprod Salerno Italy Univ Campania Luigi Vanvitelli Dept Woman Child & Gen & Specialized Surg Naples Italy Inst Genet Res IRG I-80143 Naples Italy Merck Serono SpA Global Clin Dev Rome Italy Merck KGaA Darmstadt Germany KU Leuven Univ Leuven Dept Dev & Regenerat Biomed Sci Grp Leuven Belgium

Purpose The paper aims to investigate the biological role of microRNAs secreted by preimplantation embryo into the blastocoel fluid and to detect a distinctive molecular signature for identifying embryos with the highest implantation potential. Methods We carried on a multicenter retrospective study involving five European IVF centers. We collected 112 blastocoel fluid samples from embryos on day 5 post-fertilization, cultured individually, along with data on blastocyst grade and embryo transfer outcomes. Using a custom TLDA Array, we compared the expression levels of 89 miRNAs between 33 fluids from high-quality implanted embryos and 30 fluids from high-quality not-implanted embryos. Expression differences were assessed using SAM and t-test. Additionally, correlation and function enrichment analysis and network construction were conducted to identify the biological roles of deregulated microRNAs. Results We identified six up-regulated microRNAs in the blastocoel fluid from implanted embryos, significantly and positively correlated across all samples (r >= 0.7;P <= 0.05). They could take part in pluripotency circuits, regulating and being regulated by transcription factors associated with stemness, cell growth, and embryo development. The ROC curve analysis confirmed the potential of these miRNAs as implantation classifiers. Conclusion The six miRNAs up-regulated in blastocoel fluid from implanted embryos may represent a functional molecular signature for evaluating blastocyst quality and identifying the most competent embryos. Their evaluation associated with non-invasive preimplantation genetic testing, integrating epigenetic and genomic analyses, could enhance implantation grade and allow for identification of the euploid embryo not able to implant.

关键词： blastocoel fluid Embryo development Embryo implantation Human blastocyst MicroRNAs

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Correlation among blastocoel fluid DNA level, apoptotic genes expression and preimplantation aneuploidy

REPRODUCTION AND FERTILITY

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REPRODUCTION AND FERTILITY 2025年第1期6卷 RAF-24页

作者： Khajehoseinie, Fattaneh Noormohammadie, Zahra Eftekhari-Yazdie, Poopak Gourabie, Hamid Pazhoomande, Reza Hosseinishenatale, Shirzad Bazrgare, Masood Islamic Azad Univ Dept Biol Sci & Res Branch Tehran Iran ACECR Royan Inst Reprod Biomed Reprod Biomed Res Ctr Dept Embryol Tehran Iran ACECR Royan Inst Reprod Biomed Reprod Biomed Res Ctr Dept Genet Tehran Iran Shiraz Fertil Ctr Med Genet Lab Shiraz Iran Shiraz Fertil Ctr Dept Embryol Shiraz Iran

It is believed that aneuploid embryos release cell-free DNA (cfDNA) into the blastocyst cavity during the self-correction process through the apoptotic mechanism. This study aimed to develop less invasive methods for predicting ploidy status by investigating how ploidy status affects blastocoel fluid DNA (BF-DNA) levels and apoptotic gene expression as indicators of embryo viability. Human blastocysts were classified into three groups;survivable embryo (SE), fatal single and double aneuploidy (FSDA) and multiple aneuploidy (MA) using array comparative genomic hybridization (array-CGH) by trophectoderm biopsy. Following BF aspiration and whole genome amplification, BF-DNA level was quantified. Apoptotic activity was assessed by measuring the genes TNFRSF10B, CASP2, BAX and CASP3 using real-time quantitative PCR. Day-5 intracytoplasmic sperm injection blastocysts were scored according to the Gardner and Schoolcraft system. BF-DNA levels were significantly higher in the MA vs SE group (P = 0.01), while these were not statistically significant differences between the MA and FSDA groups or between the FSDA and SE groups, P = 0.17 and P = 0.38, respectively. TNFRSF10B, CASP2 and CASP3 were overexpressed in the MA and FSDA groups compared to SE, while BAX was downregulated. We found a significant correlation between the amount of BF-DNA and apoptosis marker genes. No significant correlation was found between embryo morphology score and BF-DNA level, BF volume or apoptosis marker gene expression levels. We observed that the correlation between apoptotic activity and BF-DNA levels is influenced by the embryo's ploidy status. These findings suggest that BF-DNA level evaluation can be applicable in selecting viable embryos for transfer.

关键词： blastocoel fluid aneuploidy apoptosis cell-free DNA gene expression

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blastocoel fluid from differentiated blastocysts harbors embryonic genomic material capable of a whole-genome deoxyribonucleic acid amplification and comprehensive chromosome microarray analysis

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FERTILITY AND STERILITY 2015年第2期104卷 418-425页

作者： Tobler, Kyle J. Zhao, Yulian Ross, Ric Benner, Andy T. Xu, Xin Du, Luke Broman, Kathleen Thrift, Kim Brezina, Paul R. Kearns, William G. Johns Hopkins Univ Sch Med Dept Gynecol & Obstet Div Reprod Endocrinol & Infertil Baltimore MD 21205 USA US Army Womack Med Ctr Dept Obstet & Gynecol Ft Bragg NC 28307 USA Ft Worth Fertil Ft Worth TX USA AdvaGenix LLC Rockville MD USA Vanderbilt Univ Sch Med Dept Gynecol & Obstet Div Reprod Endocrinol & Infertil Memphis TN USA St Jude Childrens Res Hosp Dept Surg Memphis TN 38105 USA Fertil Associates Memphis Memphis TN USA

Objective: To obtain embryonic molecular karyotypes from genomic DNA (deoxyribonucleic acid) isolated from blastocoel fluid (BF) and to compare these karyotypes with the karyotypes from the remaining inner cell mass (ICM) and trophectoderm (TE) of the blastocyst. Design: Prospective cohort study. Setting: Academic center and preimplantation genetics laboratory. Patient(s): Ninety-six donated cryopreserved embryos. Intervention(s): Embryo biopsy, BF aspiration, DNA analysis using a comparative genomic hybridization microarray (aCGH). Main Outcome Measure(s): The aCGH of a single blastomere, BF-DNA, and ICM-TE. Result(s): The BF-DNA samples resulted in a successful aCGH in 63% of cases. Discordance in karyotypes was found between the BF-DNA and the ICM-TE in 52% of cases. A total of 70% of aneusomic (mosaicism), cleavage-stage embryos differentiated into euploid blastocysts. Probabilities for diagnostic accuracy were calculated and demonstrated the following: sensitivity of 0.88 (95% confidence interval [CI]: 0.62-0.98);specificity of 0.55 (95% CI: 0.39-0.70);positive predictive value of 0.41 (95% CI: 0.25-0.60);negative predictive value of 0.92 (95% CI: 0.75-0.99). Conclusion(s): Genomic DNA from the BF can be amplified and characterized by comprehensive chromosome microarrays. The results demonstrated that aneusomic cleavage-stage embryos differentiated into euploid blastocysts, possibly using a mechanism that marginalizes aneuploid nuclei into the blastocoel cavity. In addition, owing to the high discordance between the karyotypes obtained from the BF-DNA and the ICM-TE, using BF-DNA for preimplantation genetic testing is not yet advised. (C) 2015 byAmerican Society for Reproductive Medicine.

关键词： blastocoel fluid preimplantation genetic testing microarray comparative genomic hybridization preimplantation genetic screening

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blastocoel fluid aspiration improves vitrification outcomes and produces similar sexing results of in vitro-produced cattle embryos compared to microblade biopsy

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THERIOGENOLOGY 2024年 218卷 142-152页

作者： Martinez-Rodero, Iris Salas-Huetos, Albert Diaz-Munoz, Judith Ordonez-Leon, Erika Alina Garcia-Martinez, Tania Yeste, Marc Hidalgo, Carlos Olegario Mogas, Teresa Autonomous Univ Barcelona Dept Anim Med & Surg ES-08193 Cerdanyola Del Valles Spain Univ Girona Inst Food & Agr Technol Dept Biol ES-17003 Girona Spain Inst Hlth Carlos III Ctr Invest Biomed Red Fisiopatol Obes & Nutr CIBER ES-28029 Madrid Spain Harvard Univ Harvard TH Chan Sch Publ Hlth Dept Nutr Boston MA 02115 USA Brasuca Vitro Villahermosa MX-86040 Mexico Catalan Inst Res & Adv Studies ICREA ES-08010 Barcelona Spain Dept Anim Select & Reprod SERIDA ES-33394 Gijon Spain

The potential applications of in vitro-produced (IVP) cattle embryos are significantly enhanced when combined with genotype selection and cryopreservation techniques. While trophectoderm (TE) biopsies are frequently used for genotyping, cell-free DNA (cfDNA) found in blastocoele fluid (BF) arises as a less-invasive method. Moreover, the blastocoel collapse produced by BF aspiration could be beneficial for embryo cryotolerance. This study was conducted to test the BF as a source of cell free-DNA (cfDNA) and to compare the BF to the TE biopsy in terms of sexing efficiency/accuracy, embryo survival and gene expression after vitrification/warming. IVP day 7 expanded blastocysts were artificially collapsed by aspiration of BF (VIT-Collapsed) or biopsied (VIT-Biopsied). After sample collection, embryos were vitrified/warmed by the Cryotop method and individually cultured in vitro. Intact fresh non-vitrified and vitrified/warmed blastocysts served as Fresh Control and VIT-Control, respectively. After sex identification of BF or TE biopsies and the corresponding surviving embryos, amplification efficiency and sexing accuracy were assessed. There were no differences between the BF and TE biopsy samples in terms of sexing accuracy or efficiency. Although all vitrified groups showed lower post-warming reexpansion rates (p < 0.05), the blastocyst re-expansion rates in the VIT-Collapsed group were comparable to those in the Fresh Control group whereas biopsied blastocysts showed the lowest (p < 0.05) re-expansion rates. VIT-Collapsed blastocysts had hatching rates that were comparable to those of Fresh Control blastocysts but significantly higher than those of the other vitrification treatments. Proapoptotic gene BAX was overexpressed in VIT-Biopsied embryos, whereas BCL2 transcripts were more abundant in the VIT-Collapsed group. On the other hand, VIT-Biopsied embryos showed altered ATP1B1- and AQP3-mRNA levels. The analysis of the cfDNA present in the BF is an efficient, minim

关键词： Cell -free DNA blastocoel collapse blastocoel fluid Trophectoderm biopsy Microblade ICSI pipette BRY4a SAT1 PCR RT-qPCR

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Proteomic profile of maternal-aged blastocoel fluid suggests a novel role for ubiquitin system in blastocyst quality

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JOURNAL OF ASSISTED REPRODUCTION AND GENETICS 2017年第2期34卷 225-238页

作者： Tedeschi, Gabriella Albani, Elena Borroni, Elena Monica Parini, Valentina Brucculeri, Anna Maria Maffioli, Elisa Negri, Armando Nonnis, Simona Maccarrone, Mauro Levi-Setti, Paolo Emanuele Univ Milan Dipartimento Med Vet Milan Italy Fdn Filarete I-20139 Milan Italy Humanitas Res Hosp Div Gynecol & Reprod Med Humanitas Fertil Ctr Dept Gynecol Via Manzoni 56 I-20089 Milan Italy Univ Milan Dipartimento Biotecnol Med & Med Traslaz Milan Italy Univ Rome Dept Med Campus Biomed UnivVia Alvaro Portillo 21 I-00128 Rome Italy

Purpose The etiology of maternal aging, a common cause of female factor infertility and a rate-limiting step in vitro fertilization (IVF) success, remains still unclear. Proteomic changes responsible for the impaired successful pregnancy outcome after IVF with aged blastocysts have not been yet evaluated. The objective of this prospective study was to employ proteomic techniques and bioinformatic tools to enlight differences at the protein level in blastocoel fluid of aged and younger woman. Methods Protein composition of human blastocoel fluid isolated by micromanipulation from 46 blastocysts of women aged < 37 years (group A) and 29 of women aged >= 37 years (group B) have been identified by a shotgun proteomic approach based on high-resolution nano-liquid chromatography electrospray-ionization-tandem mass spectrometry (nLC-ESI- MS/MS) using label free for the relative quantification of their expression levels. Results The proteomic analysis leads to the identification and quantification of 148 proteins;132 and 116 proteins were identified in groups A and B, respectively. Interestingly, the identified proteins are mainly involved in processes aimed at fine tuning embryo implantation and development. Among the 100 proteins commonly expressed in both groups, 17 proteins are upregulated and 44 downregulated in group B compared to group A. Overall, the analysis identified 33 proteins, which were increased or present only in B while 76 were decreased in B or present only in A. Conclusions Data revealed that maternal aging mainly affects blastocyst survival and implantation through unbalancing the equilibrium of the ubiquitin system known to play a crucial role in fine-tuning several aspects required to ensure successful pregnancy outcome.

关键词： blastocoel fluid Aging Embryo implantation Proteomics Shotgun

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Failure to detect DNA in blastocoel fluid is associated with a higher live birth rate in both PGT-A and conventional IVF/ICSI cycles

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HUMAN REPRODUCTION 2023年第7期38卷 1268-1276页

作者： Gianaroli, L. Perruzza, D. Albanese, C. Azzena, S. Tabanelli, C. Ferraretti, Anna P. Magli, M. Cristina SISMeR Soc Italiana Studi Med Riprod Reprod Med Unit Bologna Italy SISMeR Soc Italiana Studi Med Riprod Reprod Med Unit Via Mazzini 12 I-40138 Bologna Italy

STUDY QUESTION Is the presence of DNA in the blastocoel fluid (BF) of expanded blastocysts, assessed by whole genome amplification (WGA), associated with the clinical outcome at the first transfer? SUMMARY ANSWER At the first transfer, blastocysts with negative BF-WGA have more chance to implant and to develop to term than those with positive BF-WGA results, both in preimplantation genetic testing for aneuploidies (PGT-A) cycles (where only euploid blastocysts resulting from the chromosomal analysis of trophectoderm (TE) biopsies were transferred) and in IVF/ICSI conventional cycles. WHAT IS KNOWN ALREADY Retrospective studies conducted in patients undergoing PGT-A have shown that the incidence of negative BF-WGA was significantly higher in TE-euploid blastocysts than in TE-aneuploid blastocysts. In addition, after the transfer of TE-euploid blastocysts, the ongoing clinical pregnancy rate was significantly higher in the group with negative BF-WGA compared with those with positive BF-WGA. STUDY DESIGN, SIZE, DURATION A prospective cohort study including 102 consecutive PGT-A patients (Group 1) and 88 consecutive conventional IVF/ICSI patients (Group 2), was conducted between January 2019 and December 2021. PARTICIPANTS/MATERIALS, SETTING, METHODS In both groups, BFs were collected from expanded blastocysts of high grade and processed for WGA. DNA amplification was evaluated by agarose gel electrophoresis for the presence (positive BF-WGA) or absence (negative BF-WGA) of a band. Directly after the BF retrieval, blastocysts from Group 1 underwent TE biopsy and vitrification. In Group 2, blastocysts were vitrified immediately after BF collection. In Group 1, only euploid blastocysts were considered for transfer according to the results of TE biopsies. In both groups, the selection of the blastocyst to be transferred was based on BF-WGA results giving priority, if available, to those with negative amplification. The primary outcome investigated was the live birth rate (

关键词： blastocoel fluid blastocysts embryo prioritization live birth delivery rate mosaicism preimplantation genetic testing for aneuploidies

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The expression of pregnancy-associated plasma protein-A (PAPP-A) in human blastocoel fluid-conditioned media: a proof of concept study

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JOURNAL OF ASSISTED REPRODUCTION AND GENETICS 2022年第2期39卷 389-394页

作者： Kavoussi, Shahryar K. Chen, Shu-Hung Wininger, John David Lal, Arnav Roudebush, William E. Lanford, Hayes C. Esqueda, Amy S. Barsky, Maya Lebovic, Dan, I Kavoussi, Parviz K. Gilkey, Melissa S. Chen, Justin Machen, Graham L. Chosed, Renee J. Austin Fertil & Reprod Med Westlake IVF 300 Beardsley LaneBldg BSuite 200 Austin TX 78746 USA Univ South Carolina Dept Biomed Sci Sch Med Greenville Greenville SC 29605 USA

Purpose The aim of this study was to determine if pregnancy-associated plasma protein-A (PAPP-A), typically measured in maternal serum and a potential predictor of adverse maternal and fetal outcomes such as spontaneous miscarriage, pre-eclampsia, and stillbirth, is expressed in blastocoel fluid-conditioned media (BFCM) at the embryonic blastocyst stage. Design This is an in vitro study. Methods BFCM samples from trophectoderm-tested euploid blastocysts (n = 80) from in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) patients were analyzed for PAPP-A mRNA. BFCM was obtained from blastocyst stage embryos in 20 uL drops. Blastocysts underwent trophectoderm biopsy for preimplantation genetic testing for aneuploidy prior to blastocyst vitrification and BFCM collection for snap freezing. cfDNA was synthesized using BFCM collected from 80 individual euploid blastocysts. Next, real-time qPCR was performed to detect expression of PAPP-A with GAPDH for normalization of expression in each sample. Results PAPP-A mRNA was detected in 45 of 80 BFCM samples (56.3%), with varying levels of expression across samples. Conclusion Our study demonstrates the expression of PAPP-A in BFCM. To our knowledge, this is the first study to report detection of PAPP-A mRNA in BFCM. Further studies are required and underway to investigate a greater number of BFCM samples as well as the possible correlation of PAPP-A expression with pregnancy outcomes of transferred euploid blastocysts. If found to predict IVF and obstetric outcomes, PAPP-A may provide additional information along with embryonic euploidy for the selection of the optimal blastocyst for embryo transfer.

关键词： blastocoel fluid Blastocyst PAPP-A Pregnancy-associated plasma protein-A IVF

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Karyotype of the blastocoel fluid demonstrates low concordance with both trophectoderm and inner cell mass

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FERTILITY AND STERILITY 2018年第6期109卷 1127-+页

作者： Tsuiko, Olga Zhigalina, Daria, I Jatsenko, Tatjana Skryabin, Nikolay A. Kanbekova, Olga R. Artyukhova, Victoria G. Svetlakov, Anatoly, V Teearu, Katre Trosin, Aleksander Salumets, Andres Kurg, Ants Lebedev, Igor N. Univ Tartu Inst Bio & Translat Med Dept Biomed Tartu Estonia Competence Ctr Hlth Technol Tiigi 61b EE-50410 Tartu Estonia Univ Tartu Inst Mol & Cell Biol Dept Biotechnol Tartu Estonia Russian Acad Sci Natl Res Med Ctr Res Inst Med Genet Lab Mol Diagnost Tomsk Russia Natl Res Tomsk State Univ Dept Cytol & Genet Tomsk Russia Tomsk Reg Perinatal Ctr Dept Assisted Reprod Technol Tomsk Russia Krasnoyarsk Ctr Reprod Med Dept Embryol Krasnoyarsk Russia East Tallinn Cent Hosp Womens Clin Tallinn Estonia Univ Tartu Inst Clin Med Dept Obstet & Gynecol Tartu Estonia Univ Helsinki Dept Obstet & Gynecol Helsinki Finland Helsinki Univ Hosp Helsinki Finland Russian Acad Sci Tomsk Natl Res Med Ctr Res Inst Med Genet Cytogenet Lab Tomsk Russia Siberian State Med Univ Dept Med Genet Tomsk Russia

Objective: To compare the genomic profiles of blastocoel fluid (BF), inner cell mass (ICM), and trophectoderm (TE) cells derived from the same blastocyst. Design: Prospective study. Setting: Academic and in vitro fertilization units. Patient(s): Sixteen donated cryopreserved embryos at blastocyst stage. Intervention(s): BF, TE, and ICM cells were retrieved from each blastocyst for chromosome analysis by means of next-generation sequencing (NGS). Main Outcome Measure(s): Aneuploidy screening and assessment of mosaicism in BF, TE and ICM samples with subsequent comparison of genomic profiles between the three blastocyst compartments. Result(s): Out of 16 blastocysts, 10 BF samples and 14 TE and ICM samples provided reliable NGS data for comprehensive chromosome analysis. Only 40.0% of BF-DNA karyotypes were fully concordant with TE or ICM, compared with 85.7% concordance between TE and ICM. In addition, BF-DNA was burdened with mosaic aneuploidies and the total number of affected chromosomes in BF was significantly higher compared with the TE and ICM. Conclusion(s): BF-DNA can be successfully amplified and subjected to NGS, but owing to increased discordance with ICM and TE, BF does not adequately represent the status of the rest of the embryo. To overcome biologic and technical challenges associated with BF sampling and processing, blastocentesis would require improvement in both laboratory protocols and aneuploidy calling algorithms. Therefore, TE biopsy remains the most effective way to predict embryonic karyotype, and the use of BF as a single source of DNA for preimplantation genetic screening is not yet advised. (C) 2018 by American Society for Reproductive Medicine.

关键词： Blastocentesis preimplantation genetic screening mosaicism blastocoel fluid next-generation sequencing

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Apoptotic qPCR gene expression array analysis demonstrates proof-of-concept for rapid blastocoel fluid-conditioned media molecular prediction

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JOURNAL OF ASSISTED REPRODUCTION AND GENETICS 2022年第7期39卷 1515-1522页

作者： Lal, Arnav Kranyak, Allison Blalock, Jonathan Athavale, Deepti Barre, Alyssa Doran, Addison Chang, T. Arthur Robinson, Randal D. Zimmerman, Shawn Wininger, J. David Fowler, Lauren A. Roudebush, William E. Chosed, Renee J. Univ South Carolina Sch Med Greenville Dept Biomed Sci 701 Grove Rd Greenville SC 29605 USA Univ Penn Sch Arts & Sci Philadelphia PA 19104 USA Univ Texas Hlth Sci Ctr San Antonio Dept Obstet & Gynecol San Antonio TX 78229 USA VIOS Fertil Inst Swansea IL 62226 USA Wake Forest Sch Med Dept Obstet & Gynecol Reprod Med Winston Salem NC 27101 USA

Purpose Successful identification of transcriptomic biomarkers within human IVF embryos may enhance implantation prediction and provide insights not available through conventional embryo biopsy genomic analysis. We demonstrate proof-of-concept for a methodology to assess overall embryo gene expression using qPCR with blastocoel fluid-conditioned media by examining the comparative presence of apoptotic genes. Methods blastocoel fluid-conditioned media were collected from 19 embryos (11 euploid) following trophectoderm biopsy of day-5 ICSI-IVF blastocysts. Media were assessed for apoptotic gene expression via qPCR. Statistical analysis of gene expression was conducted via Wilcoxon Signed-Ranks test (overall expression), multivariate ANOVA (functional gene groups), and chi-square test of independence (gene level). Results A significantly higher overall apoptotic gene expression within euploid versus aneuploid embryos (p = 0.001) was observed. There was significantly (p = 0.045) higher expression of pro-apoptotic genes between implanted and not implanted embryos. Pro- vs. anti-apoptotic gene expression from all euploid embryos approached significance (p = 0.053). The ploidy status-based claim is further substantiated at the gene level with significantly higher expression of BBC3 (p = 0.012) and BCL2L13 (p = 0.003) in euploid embryos compared to aneuploid embryos. Conclusions In this preliminary study, we demonstrate that (1) qualitative analysis of blastocoel fluid-conditioned media gene expression is possible, (2) global trends of expression are potentially related to clinical outcomes, and (3) gene-level expression trends exist and may be another viable metric for comparative expression between samples. The presence of statistical significance within analyses conducted with this sample size warrants a larger investigation of blastocoel fluid-conditioned media as an additional beneficial predictive tool for future IVF cases.

关键词： blastocoel fluid Apoptosis Aneuploidy qPCR Embryo selection Self-correction Diagnostic IVF

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microRNAs in the blastocoel fluid as accessible indicators of chromosomal normality

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REPRODUCTIVE BIOLOGY 2022年第4期22卷 100695-100695页

作者： Esmaeilivand, Masoumeh Fattahi, Amir Abedelahi, Ali Hamdi, Kobra Farzadi, Laya Goharitaban, Sepide Niknafs, Behrooz Tabriz Univ Med Sci Immunol Res Ctr Tabriz Iran Tabriz Univ Med Sci Fac Adv Med Sci Dept Reprod Biol Tabriz Iran Tabriz Univ Med Sci Womens Reprod Hlth Res Ctr Tabriz Iran Tabriz Univ Med Sci Fac Med Dept Anat Sci Tabriz Iran

MicroRNAs (miRNAs) derived from the pre-implantation blastocoel fluid (BF) have attracted interest as acces-sible biomarkers indicative of embryonic health in ongoing IVF cycles. Therefore, we investigated expression levels of some aneuploidy-associated miRNAs and implantation-related mRNAs as predictive markers for embryo chromosomal normality. In this study, the BF of 25 blastocysts that had been checked for aneuploidy (aneuploid=17 and euploid=8) was aspirated and the expression of 10 miRNAs (miR-20a, miR-30c, miR-661, miR-372, miR-142, miR-191, miR-345, miR-339, miR-141, and miR-27b) and four genes (ERBB4, SELL, ITGB3, ITGAV) were evaluated using real time-PCR. Results showed that the levels of miR-661 and miR-20a were significantly higher in the BF of the aneuploid embryos compared to the euploid group (p = 0.0017 and 0.004, respectively). A comparison of the mRNA levels between the aneuploid and euploid groups also demonstrated a significant difference in ITGAV (p = 0.013) and SELL (p = 0.0317) levels. In the euploid group, a negative correlation was found between ITGB3 and miR-30c (r =-0.71, p = 0.08), and in the aneuploid group, a positive correlation was found between ERBB4 and miR-345 (r = 0.71, p = 0.02). It can be suggested that miR-20a, miR-661, and ITGAV levels of BF could be used as less-invasive biomarkers to evaluate embryonic health. Moreover, aneuploidy-related miRNA levels were associated with levels of genes involved in embryo implantation.

关键词： blastocoel fluid Biomarker Implantation In vitro fertilization MiRNAs

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