Publications by authors named "Marefat Ghaffari Novin"

68 Publications

Meiosis Resumption of Immature Human Oocytes following Treatment with Calcium Ionophore .

Cell J 2021 Apr 1;23(1):109-118. Epub 2021 Mar 1.

Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Objective: maturation (IVM) of human oocytes is used to induce meiosis progression in immature retrieved oocytes. Calcium (Ca) has a central role in oocyte physiology. Passage through meiosis phase to another phase is controlled by increasing intracellular Ca. Therefore, the current research was conducted to evaluate the role of calcium ionophore (CI) on human oocyte IVM.

Materials And Methods: In this clinical trial study, immature human oocytes were obtained from 216 intracytoplasmic sperm injection (ICSI) cycles. After ovarian stimulation, germinal vesicle (GV) stage oocytes were collected and categorized into two groups: with and without 10 μM CI treatment. Next, oocyte nuclear maturation was assessed after 24-28 hours of culture. Real-time reverse transcription polymerase chain reaction (RT-PCR) was used to assess the transcript profile of several oocyte maturation-related genes ( and ) and apoptotic-related genes (, and ). Oocyte glutathione (GSH) and reactive oxygen species (ROS) levels were assessed using Cell Tracker Blue and 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) fluorescent dye staining. Oocyte spindle configuration and chromosome alignment were analysed by immunocytochemistry.

Results: The metaphase II (MII) oocyte rate was higher in CI-treated oocytes (73.53%) compared to the control (67.43%) group, but this difference was not statistically significant (P=0.13). The mRNA expression profile of oocyte maturation-related genes ( and ) (P<0.05) and the anti-apoptotic gene was remarkably up-regulated after treatment with CI (P=0.001). The pro-apoptotic and relative expression levels did not change significantly. The CI-treated oocyte cytoplasm had significantly higher GSH and lower ROS (P<0.05). There was no statistically significant difference in meiotic spindle assembly and chromosome alignment between CI treatment and the control group oocytes.

Conclusion: The finding of the current study supports the role of CI in meiosis resumption of human oocytes. (Registration Number: IRCT20140707018381N4).
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http://dx.doi.org/10.22074/cellj.2021.7130DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7944122PMC
April 2021

Design and Microfabrication of An On-Chip Oocyte Maturation System for Reduction of Apoptosis.

Cell J 2021 Apr 1;23(1):32-39. Epub 2021 Mar 1.

Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran. Email:

Objective: In customary assisted reproductive technology (ART), oocyte culture occurs in static micro drops of Petri dishes with vast media volume; while, the condition is dynamic. In this study, we aimed to improve the maturation efficiency of mammalian oocytes by designing an optimal microchamber array to obtain the integration of oocyte trapping and maturation within a microfluidic device and evaluate the role of microfluidic culture condition in lipid peroxidation level of the culture medium, matured oocytes apoptosis, and its comparison with the conventional static system.

Materials And Methods: In this experimental research, immature oocytes were collected from ovaries of the Naval Medical Research Institute (NMRI) mice. Oocytes were randomly laid in static and dynamic (passive and active) maturation culture medium for 24 hours. The lipid peroxidation level in oocyte culture media was assessed by measuring the concentration of malondialdehyde (MDA), and the rate of apoptosis in matured oocytes was assessed by the TUNEL assay after a-24 hour maturation period.

Results: The MDA concentration in both dynamic oocyte maturation media were significantly lower than the static medium (0.003 and 0.002 vs. 0.13 μmol/L, P<0.01). Moreover, the rate of apoptosis in matured oocytes after a-24 hour maturation period was significantly lower in passive dynamic and active dynamic groups compared with the static group (16%, 15% vs. 35%, P<0.01).

Conclusion: The dynamic culture for oocyte maturation (IVM) improves the viability of IVM oocytes in comparison with the static culture condition.
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http://dx.doi.org/10.22074/cellj.2021.7056DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7944125PMC
April 2021

Evaluation of Expression and Phosphorylation of Progesterone Receptor in Endometrial Stromal Cells of Patients with Recurrent Implantation Failure Compared to Healthy Fertile Women.

Reprod Sci 2021 Jan 15. Epub 2021 Jan 15.

Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Recurrent implantation failure (RIF) is the repeated failure of good-quality embryos in implantation process following several assisted reproduction cycles. Disruption of the endometrial receptivity is one of the main causes of RIF. Progesterone plays a pivotal role in the endometrial receptivity through the regulation of gene expression pattern by binding to its receptors in the endometrial cells. The aim of this study was to evaluate the expression level of progesterone receptor (PR) and its phosphorylated form in the endometrial stromal cells (eSC) of RIF patients and compare it to the eSC of healthy fertile women as control group. After isolation of the eSC from biopsy samples of RIF patients and healthy fertile women and their characterization, expression levels of PR mRNA, PR protein, and phospho-Ser294 PR protein were evaluated by quantitative real-time PCR and immunofluorescence staining, respectively. The results demonstrated a significant reduction in PR mRNA expression (P < 0.01.) and phospho-Ser294 PR protein (P < 0.05) level in RIF patients compared to the control group. These data for the first time suggest that the expression of PR and its phosphorylated form are impaired in RIF patients. Therefore, designing therapeutic methods for improving PR expression status and its regulation in the endometrium of RIF patients may help in improving the final reproductive outcomes of these cases.
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http://dx.doi.org/10.1007/s43032-020-00428-8DOI Listing
January 2021

Gelatin Electrospun Mat as a Potential Co-culture System for Production of Sperm Cells from Embryonic Stem Cells.

ACS Biomater Sci Eng 2020 10 18;6(10):5823-5832. Epub 2020 Sep 18.

Cellular and Molecular Research Centre, Iran University of Medical Sciences, 14496-14535 Tehran, Iran.

Engineering of 3D substrates with maximum similarity to seminiferous tubules would help to produce functional sperm cells from stem cells. Here, we present a 3D electrospun gelatin (EG) substrate seeded with Sertoli cells and determine its potential for guided differentiation of embryonic stem cells (ESCs) toward germline cells. The EG was fabricated by electrospinning, and its morphology under SEM, as well as cytobiocompatibility for Sertoli cells and ESCs, was confirmed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and cell attachment assay. Embryoid bodies (EBs) were formed from ESCs and co-cultured with Sertoli cells, induced with BMP4 for 3 and 7 consecutive days to induce the differentiation of EBs toward germline cells. The differentiation was investigated by immunocytochemistry (ICC), flow cytometry, and RT-PCR in four experimental groups of EBs (EBs cultured in gelatin-coated cell culture plates); Scaffold/EB (EBs cultured on EG); ESCs/Ser (EBs and Sertoli cells co-cultured on gelatin-coated cell culture plates without EG); and Scaffold/EB/Ser (EBs and Sertoli cells co-cultured on EG). All experimental groups exhibited a significantly increased MVH (germline-specific marker) and decreased c-KIT (stemness marker) expression when compared with the EB group. ICC and flow cytometry revealed that Scaffold/EB/Ser had the highest level of MVH and the lowest c-KIT expression at both 3 and 7 days postdifferentiation compared with other groups. RT-PCR results showed a significant increase in the germline marker () and a significant decrease in the ESC stemness marker () in Scaffold/EB compared to the EB group. The germline markers , , , , , and were significantly increased in Scaffold/EB/Ser compared to the Scaffold/EB group. Our findings revealed that the EG scaffold can provide an excellent substrate biomimicking the micro/nanostructure of native seminiferous tubules and a platform for Sertoli cell-EB communication required for growth and differentiation of ESCs into germline cells.
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http://dx.doi.org/10.1021/acsbiomaterials.0c00893DOI Listing
October 2020

Photobiomodulation preconditioned human semen protects sperm cells against detrimental effects of cryopreservation.

Cryobiology 2021 Feb 23;98:239-244. Epub 2020 Oct 23.

Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Price Institute of Surgical Research, University of Louisville, Noveratech LLC of Louisville, Louisville, KY, USA. Electronic address:

The biological consequences of semen samples preconditioning with photobiomodulation (PBM) were studied on human sperm cells post cryopreservation. Donated semen samples were collected from 22 married men with normal sperm parameters according to World Health Organization (WHO) criteria. Included samples were divided into control and PBM-preconditioning (one session, 810 nm, diode laser, and 0.6 J/cm) groups before cryopreservation procedure. Progressive sperm motility (PSM), morphology, viability, sperm mitochondrial membrane potential(MMP), intracellular reactive oxygen species (ROS) and lipid peroxidation of sperm cells were assessed post thawing. PBM preconditioning of cryopreserved semen samples most prominently increased the PSM percentage 30 min post thawing (p = 0.000).Application of PBM before cryopreservation significantly increased the number of viable spermatozoa (p = 0.000), increased significantly the number of spermatozoa with high MMP (p = 0.004) and decreased significantly the number of spermatozoa with low MMP post-thawing(P = 0. 007)compared to control group. Cryopreserved human sperm cells with PBM preconditioning showed significant decrease in the levels of intracellular ROS (47.66 ± 2.14 versus 60.42 ± 3.16, p = 0.002) and lipid peroxidation (3.06 ± 0.13 versus 3.68 ± 0.27, p = 0.05)compared to control group. Our findings, as the first evidence, indicated that PBM-preconditioning of human semen before cryopreservation provides a real and substantial advantage. This might lead to a novel strategy in improving PBM application in the procedures of assisted reproductive technologies.
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http://dx.doi.org/10.1016/j.cryobiol.2020.09.005DOI Listing
February 2021

Effects of treatment with hydroxychloroquine on the modulation of Th17/Treg ratio and pregnancy outcomes in women with recurrent implantation failure: clinical trial.

Immunopharmacol Immunotoxicol 2020 Dec 14;42(6):632-642. Epub 2020 Nov 14.

Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Aim: The imbalance of Th17/Treg cells has been recently suggested as a new risk factors for recurrent implantation failure (RIF). Furthermore Th17/Treg cells are involved in immune regulation in peripheral blood and endometrial tissue of patients with RIF. In this research, we investigated the effects of Hydroxychloroquine (HCQ) on the level and function of Th17 and Treg cells in women with RIF. It may be possible to improve pregnancy outcomes by modulating high cytokine levels.

Methods: Women with RIF received oral HCQ ( = 60) on day 4 of the menstrual cycle and continued until day 20 of the menstrual cycle and 2 days before embryo transfer and continued until the day of the pregnancy test, for a total of 16 days in another cycle. The serum levels of IL-17 and IL-10, the expression of transcription factors related to Th17 and Treg cells and the immune-reactivity of IL-17, IL-21 as Th17 related cytokines and IL-10, TGF- β as Treg related cytokines in endometrial tissues were evaluated by ELISA, real-time PCR, and fluorescent immunohistochemistry respectively.

Treatment with HCQ down-regulated Th17 related cytokines and function and up-regulated Treg related cytokines and function significantly ( < .001). RORγt, the Th17 transcription factor, expression was down-regulated and FOXP-3, the T-reg transcription factor, expression was up-regulated. The biochemical pregnancy rate was not significantly different in RIF patients before and after treatment.

Conclusion: Our results demonstrated that the administration of HCQ in RIF women with immune cell disorders during pregnancy could affect the Th17/Treg ratio and enhance Treg and diminish Th17 responses which may be associated with successful pregnancy outcomes. However, significant difference in pregnancy outcomes was not observed in the present study.
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http://dx.doi.org/10.1080/08923973.2020.1835951DOI Listing
December 2020

Effect of vitrification on biogenesis pathway and expression of development-related microRNAs in preimplantation mouse embryos.

Cell Tissue Bank 2021 Mar 9;22(1):103-114. Epub 2020 Oct 9.

Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Vitrification of embryos has been known as the most efficient cryopreservation method in assisted reproductive technology clinics. Vitrification of preimplantation embryo might be associated with altered gene expression profile and biochemical changes of vitrified embryos. Stringent regulation of gene expression in early embryonic stages is very critical for normal development. In the present study, we investigated the effect of vitrification on the canonical miRNA biogenesis pathway, and also the expression of developmental related miRNAs, in 8-cell and blastocyst mouse embryos. Although the expression pattern of the miRNA biogenesis pathway genes differed between 8-cell and blastocyst mouse embryos, vitrification did not affect the expression level of these genes in preimplantation embryos. The expression levels of miR-21 and let-7a were significantly decreased in vitrified 8-cell embryos and fresh blastocysts when compared with fresh 8-cell embryos. The expression of Stat3 was significantly reduced in blastocysts after vitrification. The alteration in the expression pattern of miRNAs, due to their mode of action, can affect broad downstream key developmental signaling pathways. Therefore, the blastocyst stage is the preferred point for embryo vitrification as they are less susceptible to cryo-damages regarding the stability of miRNAs related to the developmental and implantation competence of embryo.
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http://dx.doi.org/10.1007/s10561-020-09870-zDOI Listing
March 2021

Pyrvinium pamoate induces in-vitro suppression of IL-6 and IL-8 produced by human endometriotic stromal cells.

Hum Exp Toxicol 2021 Apr 6;40(4):649-660. Epub 2020 Oct 6.

Department of Biology and Anatomical Sciences, 274946School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Endometriosis, a chronic inflammatory disease, is identified by the presence of endometrial tissue outside the uterus. The prevalence of this disease among reproductive-age women is almost 10-15%. High levels of IL-6 and IL-8 have been found in the peritoneal fluid (PF) of women with endometriosis and are involved in its pathogenesis. Isolated stromal cells from 12 ectopic and eutopic endometrial biopsies of women with ovarian endometrioma and also 12 endometrial biopsies of nonendometriotic controls were treated with 1.1 µM pyrvinium pamoate, a Wnt/β-catenin signaling pathway inhibitor, for 72 hrs. Before treatment, mRNA gene expression and secretion of IL-6 and IL-8 were significantly higher in ectopic (EESCs) than eutopic (EuESCs) and control (CESCs) endometrial stromal cells. After treatment, mRNA gene expression and also secretion of IL-6 and IL-8 were significantly reduced. Our Findings showed that pyrvinium pamoate suppresses the mRNA gene expression and secretion of IL-6 and IL-8 in human endometriotic stromal cells. Additional investigations on this compound are required before clinical application.
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http://dx.doi.org/10.1177/0960327120964543DOI Listing
April 2021

The effect of PEGylated iron oxide nanoparticles on sheep ovarian tissue: An ex-vivo nanosafety study.

Heliyon 2020 Sep 6;6(9):e04862. Epub 2020 Sep 6.

Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Today, nanotechnology plays an important role in our ever-continuous quest to improve the quality of human life. Because of their infinitesimal size, nanostructures can actively interact and alter cellular functions. Therefore, while the clinical benefits of nanotechnology may outweigh most of the associated risks, assessment of the cytotoxicity of nanostructures in respect to cells and tissues early in product development processes is of great significance. To the best of our knowledge, no such assessment has been performed for nanomaterials on the ovarian cortex before. Herein, silica-coated, PEGylated silica-coated, and uncoated iron oxide nanoparticles (IONP) with core diameter of 11 nm (±4.2 nm) were synthesized. The oxidative stress in cultured ovarian tissue exposed to the various IONP was subsequently assessed. The results indicate that among the four groups, uncoated IONP induce the most oxidative stress on the ovarian cortex while tissues treated with PEGylated IONP exhibit no significant change in oxidative stress.
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http://dx.doi.org/10.1016/j.heliyon.2020.e04862DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7486432PMC
September 2020

Tacrolimus Improves the Implantation Rate in Patients with Elevated Th1/2 Helper Cell Ratio and Repeated Implantation Failure (RIF).

Geburtshilfe Frauenheilkd 2020 Aug 14;80(8):851-862. Epub 2020 Aug 14.

Fundacion Instituto Valenciano de Infertilidad (FIVI), Instituto Universitario IVI (IUIVI), ISS LaFe, Valencia, Spain.

An abnormal endometrial immune response is involved in the pathogenesis of repeated implantation failure (RIF), so we investigated the effectiveness of tacrolimus treatment on the endometrium of RIF patients. Ten RIF patients with elevated T-helper 1/T-helper 2 (Th1/Th2) cell ratios were recruited into a clinical study. The expression of p53, leukemia inhibitory factor (LIF), interleukin (IL)-4, IL-10, IL-17, and interferon gamma (IFN-γ) in the endometrium of patients with and without tacrolimus treatment and the association of these factors with assisted reproductive technology (ART) outcomes were investigated. Tacrolimus significantly increased the expression of LIF, IL-10, and IL-17 and decreased the expression of IL-4, IFN-γ, and the IFN-γ/IL-10 ratio in RIF patients. Tacrolimus treatment resulted in an implantation rate of 40%, a clinical pregnancy rate of 50%, and a live birth rate of 35% in RIF patients with elevated Th1/Th2 ratios who had previously failed to become pregnant despite at least three transfers of embryos. We also found a significant positive correlation between IL-10 levels and the implantation rate. Our findings suggest that RIF patients with a higher Th1/Th2 ratio could be candidates for tacrolimus therapy and that this immunosuppressive drug could be acting through upregulation of LIF, IL-10, and IL-17.
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http://dx.doi.org/10.1055/a-1056-3148DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7428373PMC
August 2020

Photobiomodulation with 810 nm Wavelengths Improves Human Sperms' Motility and Viability .

Photobiomodul Photomed Laser Surg 2020 Apr;38(4):222-231

Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Enhanced sperm motility is necessary for the successful journey of sperm inside the female genital tract, successful fertilization, and the increased chance of pregnancy. We investigated the impact of red and near-infrared (NIR) ranges of photobiomodulation (PBM) alone and together on fresh human sperm to validate an optimized PBM protocol that would maximize sperm motility and viability . We randomly divided 30 normal human semen samples into 3 different PBM protocols (red, NIR, and red+NIR lasers). Each sample was divided into four subparts, one control group sample and three experimental group samples. Each experimental group received one of the PBM protocols (red, NIR, or red+NIR). Each protocol was adjusted to three energy densities (0.6, 1.2, and 2.4 J/cm). After exposure to the selected protocol, we determined the percentage of either viable or progressive sperm motility (PSM) and measured the DNA Fragmentation Index (DFI). The NIR and red+NIR lasers at 2.4 J/cm energy density significantly increased PSM after 60 min compared with the control groups [least significant difference (LSD) test,  = 0.023 and  = 0.04, respectively]. Samples treated with the red laser at 0.6 J/cm had significantly decreased viability compared with the control group (LSD test,  = 0.003). Samples treated with the red+NIR lasers had significantly decreased viability at 0.6 J/cm ( = 0.003), 1.2 J/cm ( = 0.001), and 2.4 J/cm ( = 0.04) energy densities when compared with the control groups. The NIR laser resulted in no significant difference in sperm viability between the control and experimental groups. At 120 min after exposure, treatment with the red+NIR and red lasers at 2.4 J/cm density significantly increased DFI compared to the control groups (LSD test,  = 0.000,  = 0.007). In this study, sperm motility, viability, and DFI data confirmed the superiority of the NIR laser at 0.6 J/cm energy density compared with the red and red+NIR PBM protocols.
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http://dx.doi.org/10.1089/photob.2019.4773DOI Listing
April 2020

Improvement of in situ Follicular Activation and Early Development in Cryopreserved Human Ovarian Cortical Tissue by Co-Culturing with Mesenchymal Stem Cells.

Cells Tissues Organs 2019;208(1-2):48-58. Epub 2020 Mar 23.

Urogenital Stem Cell Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran,

Follicular loss and tissue degeneration are great challenges in ovarian tissue culture systems. Mesenchymal stem cells (MSC) secrete a cocktail of growth factors and cytokines which supports adjacent cells and tissues. The aim of the current study was to investigate the impact of human bone marrow (hBM)-MSC, as co-culture cells, on human follicular development in ovarian cortical tissue (OCT) culture. For this purpose, warmed OCT fragments were co-cultured with hBM-MSC for 8 days and compared to monocultured OCT. During the culture period, ovarian follicle survival and development in the OCT were evaluated using histological observation, follicular developmental-related genes expression, and estradiol production. Furthermore, cell proliferation and apoptosis were assessed. The results showed that there were no significant differences in conserved ovarian follicles with a normal morphology between the two groups. However, the percentage of developing follicles, as well as follicular developmental gene expression, significantly increased in the co-culture group compared to the monoculture group. On the other hand, compared with the monoculture group, the co-culture group demonstrated a significant increase in cell proliferation, indicated by Ki67 gene expression, as well as a dramatic decrease in apoptotic cell percentage, revealed by TUNEL assay. These findings indicated that co-culturing of hBM-MSC with OCT could improve follicular activation and early follicular development in human ovarian tissue culture systems.
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http://dx.doi.org/10.1159/000506303DOI Listing
August 2020

Potential of Auraptene in Improvement of Oocyte Maturation, Fertilization Rate, and Inflammation in Polycystic Ovary Syndrome Mouse Model.

Reprod Sci 2020 09 2;27(9):1742-1751. Epub 2020 Mar 2.

Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Polycystic ovary with poor-quality oocytes has remained problematic in polycystic ovary syndrome (PCOS) patients. It is well documented that the inflammation and production of reactive oxygen species (ROS) in PCOS ovaries are significantly higher than normal voluntaries. In this study, we hypothesized that auraptene (AUR), as a coumarin derivative with anti-inflammatory properties, may be effective in improvement of oocyte maturation and fertilization rate in PCOS patients. For this purpose, PCOS model was induced in NMRI mice and confirmed by ovarian histopathology observations and hormonal assays. PCOS-induced mice were administrated with AUR (PCOS-AUR) and metformin (PCOS-MET), and their effects on inflammation, apoptosis rate, oocyte maturation, and in vitro fertilization capacity were determined and compared with those normal and PCOS animals treated with sesame oil (PCOS-sesame oil) and no treatment (PCOS). Treatment with AUR and MET decreased the inflammation and apoptosis rates in PCOS mice compared with PCOS animals with no treatment. PCOS-AUR and PCOS-MET oocytes also showed higher intracellular glutathione and lower ROS concentrations compared with PCOS mice, indicating improved oocyte maturation rate. PCOS-AUR and PCOS-MET groups showed higher percentages of expansion rate and MII stage oocytes, and lower rate of abnormal oocytes compared with PCOS with no treatment. The rate of fertilization in the oocytes isolated from PCOS-AUR and PCOS-MET groups was higher than PCOS-sesame oil and PCOS groups. Our findings suggest that AUR can be considered as a potential candidate for improvement of oocyte maturation and fertilization capacity in PCOS patients, comparable to MET.
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http://dx.doi.org/10.1007/s43032-020-00168-9DOI Listing
September 2020

Effect of 1,25(OH)2-vitamin D3 on expression and phosphorylation of progesterone receptor in cultured endometrial stromal cells of patients with repeated implantation failure.

Acta Histochem 2020 Feb 24;122(2):151489. Epub 2019 Dec 24.

Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Electronic address:

Repeated implantation failure (RIF) occurs in a condition when good quality embryos fail to implant in the endometrium following several in vitro fertilization (IVF) cycles. Suboptimal endometrial receptivity is one of the main underlying factors that causes this failure. Progesterone is the key regulator of endometrial receptivity which regulates gene expression through binding to its receptors in the endometrial stromal cells (eSC). The aim of this study was to evaluate the effect of 1,25(OH)2-vitamin D3 on progesterone receptor (PR) expression level and its phosphorylation on Ser294 residues in eSC of RIF patients and healthy fertile women. After isolation of the eSC from biopsy samples of RIF patients and healthy fertile women and their characterization, the cells were incubated with vitamin D3 and the expression level of PR mRNA, PR protein and phospho-Ser294 PR protein were evaluated after treatment. The results showed that vitamin D3 treatment increases PR mRNA and protein level and phospho-Ser294 PR protein level in the isolated eSC of both RIF patients and the control group. These results suggest that vitamin D3 may possibly play a key role during the embryo implantation process by affecting the expression pattern and regulatory modifications of the PR in the eSC.
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http://dx.doi.org/10.1016/j.acthis.2019.151489DOI Listing
February 2020

Folliculogenesis-Associated Genes Expression in Human Vitrified Ovarian Tissue after Xenotransplantation in γ-Irradiated Mice.

Cell J 2020 Oct 15;22(3):350-357. Epub 2019 Dec 15.

Department of Anatomical Sciences, Faculty of Medicine, Tarbiat Modares University, Tehran, Iran. Electronic Address:

Objective: Autograft transplantation of vitrified cortical ovarian tissue is an acceptable clinical technique for fertility preservation in women. Xenograft transplantation into animal models could be useful for evaluating the safety of human vitrified ovarian tissue. This study targeted to evaluate impact of vitrification on expression of the genes associated with folliculogenesis after xenograft transplantation of human vitrified ovarian tissue to γ-irradiated mice.

Materials And Methods: In this experimental study, ovarian biopsies were gathered from six transsexual persons. The cortical section of ovarian biopsies was separated and chopped into small pieces. These pieces were randomly divided into vitrified and non-vitrified groups. In each group some pieces were considered as non-transplanted tissues and the others were transplanted to γ-irradiated female National Medical Research Institute (NMRI) mice. Before and after two weeks of xenograft transplantation, histological assessment and evaluation of the expression of folliculogenesisassociated genes ( and ) were performed in both vitrified and non-vitrified groups.

Results: Percentage of the normal follicles and expression of the all examined genes from transplanted and nontransplanted tissue were similar in both vitrified and non-vitrified groups (P>0.05). After transplantation, the normal follicle rate was significantly decreased and among the folliculogenesis-associated genes, expression of gene was significantly increased, rather than before transplantation in vitrified and non-vitrified tissues (P<0.05).

Conclusion: The vitrification method using dimethyl solphoxide and ethylene glycol (EG) had no remarkable effect on the normal follicular rate and expression of folliculogenesis-associated genes after two weeks human ovarian tissue xenografting. In addition, transplantation process can cause a significant decrease in normal follicular rate and expression of gene.
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http://dx.doi.org/10.22074/cellj.2020.6553DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6947005PMC
October 2020

MicroRNA-based regulatory circuit involved in sperm infertility.

Andrologia 2020 Feb 24;52(1):e13453. Epub 2019 Nov 24.

Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

miRNAs (MicroRNAs), known as noncoding and important endogenous factors regulating the expression protein-coding genes, are vital regulators in each biological process. Thus, this study aims to explore the key role of four microRNAs in regulating the spermatogenesis. To conduct this experiment, 55 infertile and fertile men provided the study with the sperm and testicular tissue samples. To study the spermatozoa in terms of the morphology, Diff-Quick was applied. Then, quantitative real-time polymerase chain reaction (RT-PCR) was conducted on samples. Our data indicated that in contrast to the miR-15b, significant increasing of miR-383 and miR-122 occurred in both severe oligoasthenoteratozoospermia (SOAT) and moderate oligoasthenoteratozoospermia (MOAT) compared to normal sperm group (N). In addition, it was observed that miR-15b and miR-122 increased in patients with nonobstructive azoospermia (NOA) compared with obstructive azoospermia (OA) group. Expression levels of target genes including P53, CASPASE-9 and CYCLIN D1 underwent principle changes according to miRNAs expression level. Our finding indicated that miRNAs had essential role in the regulation of spermatogenesis, and their expression altering was associated with sperm abnormalities. Thus, microRNAs can be introduced as useful biomarkers to determine male infertility reasons to choose the effective treatment.
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http://dx.doi.org/10.1111/and.13453DOI Listing
February 2020

The antioxidant curcumin postpones ovarian aging in young and middle-aged mice.

Reprod Fertil Dev 2020 Feb;32(3):292-303

Cellular and Molecular Biology Research Centre, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Postal code: 1985717443; and Corresponding authors. Emails:

Reproductive senescence is accompanied by a reduced number and quality of ovarian follicles in response to the accumulation of free radicals and the process of apoptosis. Having selected mice as models, we examined the hypothesis that curcumin as an antioxidant and anti-inflammatory agent might prevent or retard ovarian aging. Female NMRI 21-day-old mice were divided into control, vehicle and curcumin groups. In the treatment group the mice received curcumin at 100mgkg-1day-1 intraperitoneally. After 6, 12 and 33 weeks several parameters were examined including ovarian reserve, oocyte quality, oxidative status, invitro fertilisation and expression of ovulation-related (growth differentiation factor 9 (GDF-9) and bone morphogenetic protein 15 (BMP-15)) and anti-aging-related (sirtuin 1 (SIRT-1) and SIRT-3) genes. Curcumin treatment up to 12 and 33 weeks resulted in increased ovarian volume and number of follicles and was associated with elevated anti-Müllerian hormone and oestrogen and diminished FSH serum levels. Furthermore, enhanced oocyte maturation, fertilisation and embryo development plus reduced oxidative stress were seen in the curcumin group. Also, the expression of GDF-9, BMP-15, SIRT-1 and SIRT-3 genes was increased in the curcumin group. Concerning gestational age, the findings of the study suggested that administration of curcumin could delay the process of oocyte aging in a mouse model.
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http://dx.doi.org/10.1071/RD18472DOI Listing
February 2020

Does in vitro fertilization affect the expression of miRNAs and their biogenesis pathway in preimplantation mouse embryos?

Birth Defects Res 2020 01 14;112(1):62-70. Epub 2019 Oct 14.

Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Background: In vitro fertilization (IVF) is a well-accepted procedure which has been utilized for the treatment of infertile patients. As embryos at early stages of development are very vulnerable, the IVF conditions may influence genetic and epigenetic regulation of preimplantation mouse embryo.

Methods: We assessed the effect of IVF on the expression of developmental and implantation related miRNAs (miR-21, miR-93, miR-24, and let-7a), their common presumptive target (Stat3), and miRNA biogenesis pathway genes (Drosha, Dgcr8, Exportin-5, Dicer, and Ago2). in vivo 8-cell and blastocysts were compared to IVF embryos. Expression levels of miRNAs, Stat3, and miRNA biogenesis pathway genes were evaluated by qRT-PCR in in vivo (n = 8) and IVF (n = 4) embryos.

Results: The expression levels of let-7a and Stat3 were significantly reduced in IVF blastocyst when compared with in vivo (p = .004 and p = .009, respectively). Nevertheless, the IVF procedure did not influence the expression levels of miRNA biogenesis pathway components in 8-cell and blastocyst embryos.

Conclusions: Downregulation of let-7a and developmental related transcription factor, Stat3, in IVF mouse blastocysts may affect preimplantation development and implantation of embryos. Moreover, the genes of the miRNA biogenesis pathway were not changed in preimplantation mouse embryos through the IVF procedure.
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http://dx.doi.org/10.1002/bdr2.1599DOI Listing
January 2020

Expression of miR-302 in human embryo derived from in-vitro matured oocyte.

Int J Reprod Biomed 2019 Jun 29;17(6):405-412. Epub 2019 Jul 29.

Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Background: The expression of miR-302 over the period of early embryogenesis could possibly regulate the maternal transcript clearance. Zygotic transcription activation is mostly related to maternal messages degradation.

Objective: In this study, the effects of in-vitro maturation technique (IVM) on the expression of miR-302 in human embryo produced from immature and mature human oocytes (matured in vitro and in vivo, before sperm exposure) obtained from females under gonadotrophin therapy were evaluated for assisted reproduction.

Materials And Methods: Immature oocytes were cultured in vitro. The injection of oocytes-producing polar bodies was given using fresh sperm. Then, the embryo quality score was assessed in the IVM group compared with the control group. In both the groups, embryos with normal morphology were included in the molecular study. Only one blastomere was removed from three-day embryos and then the embryos were frozen. The expression of miR-302 in embryos was measured through quantitative real-time polymerase chain reaction.

Results: Our data showed a significant reduction of miR-302 expression in the IVM group vs. the control group (p = 0.02). The embryo quality score showed a significant difference between the two groups (p = 0.01).

Conclusion: The present study demonstrated that the IVM process had a negative effect on the expression level of miR-302 in human pre-implantation embryos. Considering the major role of expression miR-302, a reduced potential in miR-302 expression could be related to a decrease in the early embryonic development.
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http://dx.doi.org/10.18502/ijrm.v17i6.4812DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6719516PMC
June 2019

Thymoquinone reduces intracytoplasmic oxidative stress and improves epigenetic modification in polycystic ovary syndrome mice oocytes, during in-vitro maturation.

Mol Reprod Dev 2019 08 17;86(8):1053-1066. Epub 2019 Jun 17.

Department of Pharmacology, School of Medicine and Neuroscience Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Although in-vitro maturation (IVM) of oocytes has been presented as an alternative treatment to traditional stimulated in-vitro fertilization, the culture condition can be improved by natural antioxidants. Thus, we investigated the protective effect of Thymoquinone (TQ) during IVM in the polycystic ovary syndrome (PCOS) mice model. The induction of PCOS was made by dehydroepiandrosterone via subcutaneous injection, in prepubertal female B6D2F1-mice. After 21 days later, germinal vesicle (GV)-stage-oocytes were extracted and incubated in IVM media containing 0, 1.0, 10.0, and 100.0 μM of TQ. To assess fertilization and blastulation rates, after 22-24 hr, the treated oocytes were fertilized in-vitro with epididymal spermatozoa. Some other oocytes were evaluated for maturation, epigenetic, and oxidative stress markers. Similarly, the mRNA expression of epigenetic enzymes genes (Dnmt1 and Hdac1), three maternally derived genes (Mapk, CyclinB, and Cdk1) and apoptosis-related genes (Bax and Bcl2) were assessed. Our results showed that the maturation, fertilization, and blastulation rates were significantly higher in the 10.0 μM TQ-treated group compared with the untreated group and likewise with in-vivo matured oocytes. The Bax expression was reduced in 10.0 μM TQ matured oocytes, but Bcl2, Dnmt1, Hdac1, Cdk1, and Mapk were upregulated in this group compared to other groups. Furthermore, dimethylation of histone-3 at lysine-9 (H3K9m2) and DNA methylation were significantly increased whereas H4K12 acetylation (H4K12ac) was decreased in the 10.0 μM TQ-treated group in comparison with control and in-vivo matured oocytes. Therefore, our results are suggesting that 10.0 μM TQ may enhance the developmental competence of PCOS oocytes via the modulation of oxidative stress and epigenetic alterations.
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http://dx.doi.org/10.1002/mrd.23222DOI Listing
August 2019

Serum and Peritoneal Fluid Cytokine Profiles in Infertile Women with Endometriosis.

Iran J Immunol 2019 Jun;16(2):151-162

Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran,Iran.

Background: Endometriosis is a chronic inflammatory disease with the growth of endometrial cells out of uterus and in the peritoneal cavity. T cell subsets participate in the establishment and progress of the disease by producing different cytokines.

Objective: To investigate a group of cytokines related to Th1/Th2/Th17/Treg subsets within both peripheral blood and peritoneal fluid (PF) samples from infertile endometriosis women.

Methods: Peripheral blood and PF samples were collected from 30 infertile endometriosis and 30 non-endometriosis fertile women during laparoscopy. Concentration of cytokines, including TNF-α, IFN-γ, TGF-β1, IL-4, IL-10, IL-17 and IL-23 were evaluated using ELISA method.

Results: Results indicated that the concentration of IFN-γ within serum was significantly reduced in endometriosis group (p=0.001). Regarding PF cytokines, TGF-β1 was increased in endometriosis group (p=0.030). Furthermore, the ratios of IFN-γ/TGF-β1 and IL-17/IL-23 were significantly different between endometriosis and non-endometriosis women in serum samples (p<0.001 and p<0.01 respectively). The ratios of TNF-α/IL-10 and IL-17/IL-10 were also significantly different regarding PF samples between the two studied groups (p<0.04 and p<0.03 respectively). Finally, significant correlations were observed between the levels of IL-17 and IL-23, inflammatory and anti-inflammatory cytokines, in both samples and serum to PF inflammatory cytokines.

Conclusion: Based on the results of the present study, in women with endometriosis, the disturbance of cytokines network might gradually activate the inflammatory responses and tissue repair, resulting in endometriosis development after several years.
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http://dx.doi.org/10.22034/IJI.2019.80258DOI Listing
June 2019

Protective effect of gallic acid on apoptosis of sperm and in vitro fertilization in adult male mice treated with cyclophosphamide.

J Cell Biochem 2019 10 28;120(10):17250-17257. Epub 2019 May 28.

Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Background: Alteration of free radicals (reactive oxygen species) causes mammals' sperm damage. Gallic acid (GA) is known as an antioxidant which is effective against oxidative stress. The purpose of this study was to evaluate the antioxidant effects of GA on the sperm apoptosis and in vitro fertilization (IVF) in adult male mice treated with cyclophosphamide (CP).

Materials And Methods: Following a pilot study to find the dose responses of GA, 40 adult male naval medical research institute (NMRI) mice (32 ± 3 g) were divided into five groups (n = 8): control, sham (normal saline, NS: 0.2 mL per day), CP (15 mg kg per week; intraperitoneal, IP), GA (12.5 mg kg per day; IP), and GA+CP. After the treatment, sperm parameters were analyzed. The apoptosis of sperm was measured by Annexin-PI staining method followed by flow cytometry detection. Fertility was assessed by IVF method among the groups.

Results: The difference in sperm parameter and fertility rate between the control (% 80.05 ± 6.53) and cyclophosphomide groups (% 51.82 ± 10.78) was significant (P < .001) but GA plus CP (% 78.16 ± 5.71) restored the fertilization rate (P < .001). Also, a remarkable increase was noted regarding apoptotic sperm in CP group vs the control group. The comparison in the five groups shows that GA cotreatment was significantly effective in reducing the apoptosis rate caused by cyclophosphamide (P < .05).

Conclusion: It was ultimately attained that GA has a potent antioxidant effect which could inhibit the detrimental effect of CP on the apoptosis and fertility rate of sperm in the mouse.
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http://dx.doi.org/10.1002/jcb.28987DOI Listing
October 2019

Differentiation of spermatogonial stem cells by soft agar three-dimensional culture system.

Artif Cells Nanomed Biotechnol 2019 Dec;47(1):1772-1781

a Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences , Tehran , Iran.

Since proliferation and differentiation of spermatogonial stem cells (SSCs) in culture system provide successful transplantation in this study, culture of human SSCs was compared to SACS (soft agar culture system), gelatin and control groups. The cells were isolated from seminiferous tubules of non-azoospermia patients (NOA) and cultured in DMEM for 3 weeks. The presence of SSCs in culture system was confirmed by immunocytochemistry of GFR-α1 and ITGα6 antibodies. The proliferated cells were cultured in three mentioned groups in the presence of retinoic acid and Sertoli cells conditioned medium for another 2 weeks. The number of colonies in the SACS group was significantly higher than two other groups. Before 2 weeks of culture, only Oct4 expression was observed in testicular cells (2.32 ± 0.25). After 2 weeks, the expression of Oct4 in the gelatin group was higher than that of the SACS group on day 7. The maximum expression of Stra8 was observed in SACS and gelatin groups after 7 days, but its expression was significantly decreased after 14 days of culture ( < .05). The expression of Scp3 and Acrosin genes were higher after 14 days in the SACS group compared to other groups. SACS has positive effects on proliferation and differentiation of hSSCs.
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http://dx.doi.org/10.1080/21691401.2019.1575230DOI Listing
December 2019

Three-dimensional electrospun gelatin scaffold coseeded with embryonic stem cells and sertoli cells: A promising substrate for in vitro coculture system.

J Cell Biochem 2019 08 11;120(8):12508-12518. Epub 2019 Apr 11.

Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

In this study, we present an electrospun gelatin (EG) scaffold to mimic the extracellular matrix of the testis. The EG scaffold was synthesized by electrospinning and crosslinked with glutaraldehyde vapor to decrease its water solubility and degradation rate. The scanning electron microscope micrographs showed the homogenous morphology of randomly aligned gelatin fibers. The average diameter of gelatin fibers before and after crosslinking was approximately 180 and 220 nm, respectively. Modulus, tensile strength, and elongation at break values were as 161.8 ± 24.4 MPa, 4.21 ± 0.54 MPa, and 7.06 ± 2.12 MPa, respectively. The crosslinked EG showed 75.2% ± 4.5% weight loss after 14 days with no changes in the pH value of degradation solution. Cytobiocompatibility of the EG for sertoli cells and embryonic stem cells (ESCs) was determined in vitro. Sertoli cells were isolated from mouse testis and characterized by immunostaining and flow cytometry. The effects of EG on proliferation and attachment of both sertoli cells and ESCs were examined. The EG scaffolds exhibited no cytotoxicity for sertoli and ESCs. Both sertoli and ESCs were well attached and grown on EG. Coculture of sertoli and ESCs on EG showed better ESCs adhesion compared with ESCs alone. Our findings indicate the potential of EG as a substrate for proliferation, adhesion, and coculture of sertoli and ESCs and may be considered as a promising engineered microenvironment for in vitro coculture system with the aim of guiding stem cells differentiation toward sperm-producing cells.
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http://dx.doi.org/10.1002/jcb.28517DOI Listing
August 2019

Protective Effect of Gallic Acid on Testicular Tissue, Sperm Parameters, and DNA Fragmentation against Toxicity Induced by Cyclophosphamide in Adult NMRI Mice.

Urol J 2020 01 26;17(1):78-85. Epub 2020 Jan 26.

Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Purpose: This study aimed to investigate the protective effect of Gallic acid (GA) on the Cyclophosphamide (CP) toxicity induced in the reproductive system.

Materials And Methods: After a pilot study for dose responses of Gallic acid ,Forty adult male NMRI mice were divided into 5 groups (n=8): control, sham (NaCl Serum: 0.2mL per day), CP (15 mg kg-1 per week; IP), GA (12.5 mg kg-1 per day ; IP) and GA (12.5 mg kg-1 per day ; IP) +CP(15 mg kg-1 per week; IP). After treatment, the left testis was detached and used for Histological examination and right testis used for Malondialdehyde (MDA) measures. Left caudal epididymis was placed in the Ham's F10 medium and released spermatozoa were used in order to analyze sperm parameters. Sperm DNA fragmentation was assessed by Sperm Chromatin Dispersion (SCD) method.

Results: In the CP group, there was a significant increase in the sperm DNA fragmentation (% 57.89 ± 23.91) compared with control group (% 24.52 ± 10.27). That was significantly improved by GA  (12.5 mg kg-1 per day ; IP)  in GA+CP group (% 28.4 ± 8.85) compared to CP group (p< .001).A significant increase was reported about MDA levels in CP group (6.26 ± 2.59) in compared with the control group (4.30 ± 2.05), But GA (3.24 ± 1.33) decreased it in GA+ CP group (p< .01).  The histopathological investigation revealed marked testicular atrophy in CP group, whereas GA diminished these deviations (P< .05).

Conclusion: Gallic acid can modify the reproductive toxicity of cyclophosphamide in NMRI mice and increase the antioxidant capacity of testis tissue.
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http://dx.doi.org/10.22037/uj.v0i0.4858DOI Listing
January 2020

Effect of Artificial Oocyte Activation on Intra-Cytoplasmic Sperm Injection Outcomes in Patients with Lower Percentage of Sperm Containing Phospholipase Cζ: A Randomized Clinical Trial.

J Reprod Infertil 2019 Jan-Mar;20(1):3-9

Infertility and Reproductive Health Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Background: Artificial oocyte activation (AOA) is a specialized method in assisted reproductive technique (ART). According to increasing concern about using AOA, it is necessary to evaluate sperm-borne oocyte activating factors (SOAFs) including phospholipase C zeta (PLCζ). In this study, PLCζ before AOA was evaluated first and then the impact of AOA on pre-implantation embryo development was investigated.

Methods: This prospective clinical trial enrolled couples subjected to ICSI. By evaluating PLCζ, semen samples were categorized into two groups; I (Control) and II (PLCζ deficient). Retrieved oocytes from partners were put into three categories: control group (Injected with sperm from group I, n=113), group without AOA (Injected with sperm from group II and no exposure to AOA, n=106), and group AOA (Injected with sperm from group II and exposure to AOA, n=114). Finally, fertilization results were compared via Kruskal-Wallis followed by Dunn's multiple comparison test. The p<0.05 was considered statistically significant.

Results: Fertilization rate was significantly lower in the group without AOA compared to control group (41.9±6.3 . 78.1±4.7, p<0.001). AOA improved fertilization rate in group AOA compared to the group without AOA (69.5±3.9 . 41.9±6.3, p<0.01); however, cleavage (91.7±2.8, 90.9±4.6, and 95.2±3.4, respectively) and embryo quality (2.5±0.1, 2.3±0.2, and 2.4±0.2, respectively) scores were not substantially different between groups of control, with and without AOA.

Conclusion: We showed that PLCζ can be considered as a good biomarker in evaluation of oocyte activation capability. Further studies are required to establish the best use of PLCζ as a biomarker in clinics.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6386797PMC
March 2019

Seminal exosomes induce interleukin-6 and interleukin-8 secretion by human endometrial stromal cells.

Eur J Obstet Gynecol Reprod Biol 2019 Apr 19;235:71-76. Epub 2019 Feb 19.

Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Electronic address:

Objective: Exosomes are extracellular microvesicles that participate in intercellular communication. Seminal plasma (SP) contains very large amounts of exosomes which are deposited in female genital tract after insemination. Although the response of vaginal cells to seminal exosomes (SE) is recently being elucidated, the interaction of uterine cells with SE is still unknown. Here, we aimed to evaluate the effect of SE on cytokine secretion by human endometrial stromal cells (eSC).

Study Design: Exosomes were isolated from the semen samples of healthy men with proven fertility and characterized using common exosome characterization methods. Human eSC were isolated from endometrial biopsies obtained from healthy premenopausal women. For exosome internalization analysis, SE were labeled with PKH67 green fluorescent dye and incubated with the cells. For investigating the effect of SE on cytokine secretion of eSC, we measured levels of interleukin (IL)-6, IL-8, IL-10, IL-1α, and leukemia inhibitory factor (LIF) in the culture supernatants of control and experimental groups by enzyme-linked immunosorbent assay (ELISA) after 24 h of incubation.

Results: Our results demonstrated that SE are internalized by eSC and subsequently induce them to produce IL-6 and IL-8, the cytokines which are involved in the immunology of embryo implantation.

Conclusion: The findings of the present study suggest that SE contribute to the immunoregulatory functions of SP in the uterus and may participate in embryo implantation process. Therefore dysfunction of intracellular machineries of SE biogenesis and secretion, inadequate production, defective transportation to the uterus and impaired communication with endometrium may play a distinct role in pathophysiology of embryo implantation failure.
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http://dx.doi.org/10.1016/j.ejogrb.2019.02.010DOI Listing
April 2019

Impact of sperm DNA fragmentation on ICSI outcome and incidence of apoptosis of human pre-implantation embryos obtained from in vitro matured MII oocytes.

Biochem Biophys Res Commun 2019 02 17;510(1):110-115. Epub 2019 Jan 17.

Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Background: Sperm DNA integrity and oocyte quality significantly affect embryo development and survival. The current study evaluated embryo development and quality, as well as the expression level of apoptosis-related genes and microRNAs in embryo derived from in vitro matured MII oocytes according to sperm DNA fragmentation (SDF) level.

Methods: The semen and immature oocytes were collected from 50 ICSI cycles with any recognizable female factor infertility. After ovarian stimulation, germinal vesicle stage (GV) oocytes were collected and incubated in in vitro maturation (IVM) medium for 24 h. Next, reactive oxygen species (ROS) level of media culture was determined. Using by sperm chromatin dispersion (SCD) test, the SDF levels of processed semen were assessed and categorized into SDF ≤ 30% and SDF>30%. Seventy two hours after intracytoplasmic injection, the embryo development and quality score were recorded in the groups I (GV-MII + SDF≤ 30%) and II (GV-MII + SDF> 30%). Also, the apoptosis incidence of embryos at morula stage was evaluated at molecular and cellular levels by quantitative real time PCR and TUNEL staining, respectively.

Results: Cleavage rate did not differ between two groups. The quality score of embryos obtained from IVM matured oocytes and high level of SDF was significantly lower than that of low level of SDF (P < 0.05). The embryos from group II had a significant reduction of the expression of BCL-2 compared to group I (P < 0.05). Also, they showed an increase in relative transcription of pro-apoptotic microRNAs; miR 15a and miR 16-1 versus group I (P < 0.05). A rise of TUNEL positive blastomers of embryo was observed at group II versus group I, but it did not reach to significantly level.

Conclusion: The IVM oocytes, probably, did not suffice to recover the high level of paternal genomic damage and inhibition of apoptosis pathway beginning.
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http://dx.doi.org/10.1016/j.bbrc.2019.01.056DOI Listing
February 2019

Does blastomere removal alter the expression level of miR-Let7a and its target genes following mouse embryo biopsy?

J Cell Biochem 2019 06 2;120(6):9430-9436. Epub 2018 Dec 2.

Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Embryo manipulations may cause the misexpression of various genes, most of which play critical roles in the regulation of implantation. This study aimed to evaluate the effects of embryo biopsy on the expression of miR-Let-7a and its gene targets including ErbB4, Tgf-α, Itg-αv, Itg β3 on the implantation of mouse embryo. Embryos were produced by in vitro fertilization followed by blastomere biopsy at the eight-cell stage. The effects of blastomere removal on the expression of genes ErbB4, Tgf-α, Itg αv, Itg β3, and miR-Let-7a as well as the alteration of the blastocyst cell number were compared in both biopsied and non-biopsied groups. Finally, blastocyst attachment was assessed on culture dishes precoated with Fibronectin. The results revealed that there were no significant differences between the biopsied and non-biopsied embryos with reference to the blastocyst formation rates, the average inner cell mass, trophectoderm cell number, and percentage of attachment of blastocysts (P > 0.05). The expression of ErbB4, Itg-β3, Itg-αv, TGF-α transcripts, and miR-Let-7a in blastocysts biopsied embryos did not differ from the non-biopsied blastocysts (P > 0.05). The results demonstrated that the preimplantation embryo development and attachment of biopsied embryos in vitro is not adversely affected by one blastomere biopsy at the eight-cell stage embryo.
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http://dx.doi.org/10.1002/jcb.28218DOI Listing
June 2019