scholarly journals Cell-Free, Embryo-Specific sncRNA as a Molecular Biological Bridge between Patient Fertility and IVF Efficiency

2019 ◽  
Vol 20 (12) ◽  
pp. 2912 ◽  
Author(s):  
Angelika V. Timofeeva ◽  
Vitaliy V. Chagovets ◽  
Yulia S. Drapkina ◽  
Nataliya P. Makarova ◽  
Elena A. Kalinina ◽  
...  
Keyword(s):  
Embryo Development ◽  
Culture Media ◽  
Expression Profiles ◽  
Embryo Implantation ◽  
Human Reproduction ◽  
Vitro Fertilization ◽  
Small Noncoding Rnas ◽  
Polymerase Chain ◽  
Next Generation Sequencing Ngs

Small noncoding RNAs (sncRNAs) are key regulators of the majority of human reproduction events. Understanding their function in the context of gametogenesis and embryogenesis will allow insight into the possible causes of in vitro fertilization (IVF) implantation failure. The aim of this study was to analyze the sncRNA expression profile of the spent culture media on day 4 after fertilization and to reveal a relationship with the morphofunctional characteristics of gametes and resultant embryos, in particular, with the embryo development and implantation potential. Thereto, cell-free, embryo-specific sncRNAs were identified by next generation sequencing (NGS) and quantified by reverse transcription coupled with polymerase chain reaction (RT-PCR) in real-time. Significant differences in the expression level of let-7b-5p, let-7i-5p, piR020401, piR16735, piR19675, piR20326, and piR17716 were revealed between embryo groups of various morphological gradings. Statistically significant correlations were found between the expression profiles of piR16735 and piR020401 with the oocyte-cumulus complex number, let-7b-5p and piR020401 with metaphase II oocyte and two pronuclei embryo numbers, let-7i-5p and piR20497 with the spermatozoid count per milliliter of ejaculate, piR19675 with the percentage of linearly motile spermatozoids, let-7b-5p with the embryo development grade, and let-7i-5p with embryo implantation. According to partial least squares discriminant analysis (PLS-DA), the expression levels of let-7i-5p (Variable Importance in Projection score (VIP) = 1.6262), piR020401 (VIP = 1.45281), and piR20497 (VIP = 1.42765) have the strongest influences on the implantation outcome.

Effects of Different Types of Incubators on Embryo Development and Clinical Outcomes

2020 ◽  
Author(s):  
Ji Liu ◽  
Yan-Hua Zhou ◽  
Xiao-Xiao Wang ◽  
Ling-Xi Tong ◽  
Yan-Hong Li ◽  
...  
Keyword(s):  
Clinical Outcomes ◽  
Embryo Development ◽  
Culture Media ◽  
Controlled Trial ◽  
Formation Rate ◽  
Fertilization Rate ◽  
Vitro Fertilization ◽  
Water Jacket ◽  
Different Types

Abstract Background: Different types of incubators have been designed for gamete and embryo culture in the past few years. The main differences of these incubators are humidity, temperature and gas control system, which play important roles in regulating the steady state of culture media. The objective of this study was to compare the effects of different types of incubators (air jacket incubators and water jacket incubators) on embryo development and clinical outcomes in human in vitro fertilization (IVF).Methods: First, the physical performances of different incubators were tested by mimicking routine IVF procedures. After that, in a randomized controlled trial, 1013 cumulus oocyte complexes from 43 patients were equally divided into two groups, fertilized and cultured in two types of incubators to analyze the effects of different types of incubators on embryo development and clinical outcomes. Results: We found that temperature recovery time in the air jacket incubator was significantly shorter than that in water jacket incubator. Although the O2 recovering time was also significantly shorter in the air jacket incubator as compared with the water jacket incubator, no significant differences were observed in the CO2 recovering time between two groups, which was also verified by pH recovering time of culture media. Besides, the temperature of culture medium in the dish covered with oil recovered more quickly in the air jacket incubators than that in water jacket incubators. However, there were no significant differences observed in the fertilization rate, Day 3 high-quality embryo formation rate, blastocyst formation rate, good blastocyst rate and clinical outcomes between two groups.Conclusions: These results indicate that the microenvironment, especially the temperature, in air jacket incubator recover faster than that in conventional water jacket incubator, however, there were no significant differences in embryo development and clinical outcomes between two types of incubators.

scholarly journals Effects of Progesterone on in Vitro Developmental Competence of Bovine Embryos

2020 ◽  
Vol 8 (1) ◽  
pp. 42
Author(s):  
Orhan Örnek ◽  
Yusuf Ziya Güzey
Keyword(s):  
Embryo Development ◽  
Culture Media ◽  
Early Embryo ◽  
Developmental Competence ◽  
Direct Role ◽  
Vitro Fertilization ◽  
Morula Stage ◽  
Vitro Development

Progesterone plays a key role in the establishment and maintenance of pregnancy in mammalian. Increasing levels of circulating progesterone in the post-conception period are associated with conceptus elongation and high pregnancy rates in cattle. Contradictory results are available on the direct role of progesterone in early embryo development. The objective of this study was to evaluate direct effects of progesterone on in vitro development of cattle embryos. Immature oocytes collected from slaughtered animals and cultured in the presence of different concentrations of progesterone (25, 50, 100 ng/mL) following in vitro fertilization. Cleavage rates in 25 and 50 ng/mL concentrations of progesterone were significantly higher than those in controls and 100 ng/mL. Rate of embryos that reached to the morula stage was similar in all groups. Supplementation of 25 and 50 ng/mL progesterone to the culture media significantly increased blastocyst yield while 100 ng/mL progesterone resulted in a decrease. As a conclusion, we can suggest that progesterone supplementation in in vitro culture may support embryo development at low levels.

336 SHEEP OOCYTES MATURED IN A SIMPLEX MEDIUM (HECM-1), AN ANSWER TO IN VITRO FERTILIZATION

2006 ◽  
Vol 18 (2) ◽  
pp. 275
Author(s):  
C. Navarro-Maldonado ◽  
Y. Ducolomb-Ramirez ◽  
A. Galindo-Rodriguez ◽  
A. Rosado-Garcia
Keyword(s):  
In Vitro Fertilization ◽  
Embryo Development ◽  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Culture Media ◽  
Sperm Quality ◽  
Complete Control ◽  
Vitro Fertilization ◽  
Significant Difference

In vitro maturation and in vitro fertilization (IVM and IVF) of mammalian oocytes still show unsatisfactory results when applied to the study of embryo development. This is probably due to inadequate information about the use of media components and supplements for oocyte maturation and to a discrepancy between results obtained by focusing strictly on oocyte maturation and those that are focused on IVF. A conventional medium that provides adequate results in studies of oocyte maturation (TCM-199) contains hypoxanthine, phosphate ions, and glucose, all known to inhibit embryo development in vitro in some species. In contrast, it has been shown that a simpler medium (HECM-9) increases embryo development in bovine although its use for oocyte maturation has not been defined. This medium contains taurine (an amino acid that reduces intracellular peroxide content) and is supplemented with polyvinyl alcohol (PVA) instead of using protein components, making it a simple defined medium that reduces variability in embryo development. Adding sodium panthothenate to media also confers cell protection against reactive oxygen species. Finally, supplements such as epidermal growth factor (EGF) increase the number of oocytes that complete maturation (Metaphase II, MII) and facilitate embryo development. An adequate combination of our knowledge about in vitro maturation and fertilization of oocytes, together with the requirements for embryo development, is important for the preparation of culture media to study regulatory mechanisms for embryo development and to increase the number of viable and normal term individuals. In this study we compared the effects of HECM-9 (containing panthothenate) vs. TCM-199 (both media supplemented with PVA, EGF, and FSH/LH) on the integrated processes involving IVM and IVF. No significant differences were found between the results obtained with these media in relation to oocyte maturation (65% MII for HECM-9 vs. 71% for TCM-199); however, those oocytes matured in HECM-9 showed a highly significant difference in in vitro fertilization using a conventional IVF medium (SOFm) (25% in HECM-9 vs. 6% in TCM-199). Maturation results were relatively low but in accordance with those reported by other groups, whereas IVF results are lower than those reported in the literature, perhaps because we have been using frozen and thawed samples and do not have complete control over the sperm quality. At present, we are extending our investigation using fresh semen samples.

scholarly journals Expression of extracellular matrix metalloproteinase inducer and matrix metalloproteinases during mouse embryonic development

Reproduction ◽  
2007 ◽  
Vol 133 (2) ◽  
pp. 405-414 ◽  
Author(s):  
Li Chen ◽  
Masaaki Nakai ◽  
Robert J Belton ◽  
Romana A Nowak
Keyword(s):  
Extracellular Matrix ◽  
Matrix Metalloproteinases ◽  
Matrix Metalloproteinase ◽  
Embryo Development ◽  
Mouse Embryo ◽  
Expression Profiles ◽  
Embryo Implantation ◽  
Rt Pcr ◽  
Extracellular Matrix Metalloproteinase Inducer

Mouse embryo implantation is a highly invasive and controlled process that involves remodeling and degradation of the extracellular matrix of the uterus. Matrix metalloproteinases (MMPs) are the main proteinases facilitating this process. Extracellular matrix metalloproteinase inducer (EMMPRIN) can stimulate the production of MMPs and is required for successful implantation in the mouse. The aims of the present study were to examine the expression profiles of mRNA and proteins for EMMPRIN and MMPs in the developing mouse embryo in vitro, and to study whether EMMPRIN protein induces the production of MMPs by mouse blastocysts. EMMPRIN mRNA, detected by RT-PCR, was present at all stages of embryo development from the one-cell to the blastocyst outgrowth. EMMPRIN protein, observed by confocal microscopy, was present on the cell surface at the same stages of development as was the mRNA. Of seven MMPs studied, murine collagenase-like A (Mcol-A), murine collagenase-like B (Mcol-B) and gelatinase A (MMP-2) mRNAs were detected only in blastocyst outgrowths by RT-PCR. Gelatinase B (MMP-9) mRNA was detected both in expanded blastocysts and blastocyst outgrowths. MMP-2 and -9 proteins were detected in the cytoplasm of outgrowing trophoblast cells. Collagenase-2 (MMP-8), collagenase-3 (MMP-13), or stromelysin-1 (MMP-3) mRNAs were not present at any stage of pre- or peri-implantation mouse embryo development. Quantitative RT-PCR analyses showed that recombinant EMMPRIN protein did not stimulate MMP-2 or -9 expression by mouse blastocyst outgrowths. These data suggest that EMMPRIN may regulate physiological functions other than MMP production by mouse embryos during implantation.

scholarly journals Telomere Length in Pig Sperm Is Related to In Vitro Embryo Development Outcomes

Animals ◽  
2022 ◽  
Vol 12 (2) ◽  
pp. 204
Author(s):  
Jordi Ribas-Maynou ◽  
Yentel Mateo-Otero ◽  
Marina Sanchez-Quijada ◽  
Sandra Recuero ◽  
Ariadna Delgado-Bermúdez ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Telomere Length ◽  
Embryo Development ◽  
Sperm Quality ◽  
Quality Parameters ◽  
Human Reproduction ◽  
Vitro Fertilization ◽  
Development Outcomes ◽  
Sperm Telomere Length

Telomere length has attracted much interest as a topic of study in human reproduction; furthermore, the link between sperm telomere length and fertility outcomes has been investigated in other species. This biomarker, however, has not been much explored in other animals, such as pigs, and whether it is related to sperm quality and fertility outcomes remains unknown. The present work aimed to determine the absolute value of telomere length in pig sperm, as well as its relationship to sperm quality parameters and embryo development. Telomere length was determined through quantitative fluorescence in situ hybridization (qFISH) in 23 pig sperm samples and data were correlated to quality parameters (motility, morphology, and viability) and in vitro fertilization outcomes. We found that the mean telomere length in pig sperm was 22.1 ± 3.6 kb, which is longer than that previously described in humans. Whilst telomere length was not observed to be correlated to sperm quality variables (p > 0.05), a significant correlation between telomere length and the percentage of morulae 6 days after in vitro fertilization was observed (rs = 0.559; 95%C.I. = (−0.007 to 0.854); p = 0.047). Interestingly, this correlation was not found when percentages of early blastocysts/blastocysts (rs = 0.410; 95%C.I. = (−0.200 to 0.791); p = 0.164) and of hatching/hatched blastocysts (rs = 0.356; 95%C.I. = (− 0.260 to 0.766); p = 0.233) were considered. Through the separation of the samples into two groups by the median value, statistically significant differences between samples with shorter telomeres than the median and samples with longer telomeres than the median were found regarding development to morula (11.5 ± 3.6 vs. 21.8 ± 6.9, respectively) and to early blastocyst/blastocysts (7.6 ± 1.4 vs. 17.9 ± 12.2, respectively) (p < 0.05). In the light of these results, sperm telomere length may be a useful biomarker for embryo development in pigs, as sperm with longer telomeres lead to higher rates of morulae and blastocysts.

scholarly journals MiR-191-5p Is Upregulated in Culture Media of Implanted Human Embryo on Day Three of Development

2020 ◽  
Author(s):  
Ricardo Josue Acuña-González ◽  
Fela Vanesa Morales-Hernández ◽  
Jorge Skiold López-Canales ◽  
Jair Lozano-Cuenca ◽  
Mauricio Osorio-Caballero ◽  
...  
Keyword(s):  
Embryo Development ◽  
Human Embryo ◽  
Culture Medium ◽  
Culture Media ◽  
Expression Profiles ◽  
Uterine Cavity ◽  
Common Criteria ◽  
Significant Difference ◽  
Human Embryo Development

Abstract Background: Morphologic features are the most common criteria for selecting human embryo to be transferred to the receptive uterine cavity. However, such characteristics are not valid for embryos in cellular arrest. The aim of this study was to quantify the expression profile of hsa-miR-21-3p, -24-1-5p, -191-5p, and -372-5p on day 3 of culture media from in vitro fertilization (IVF) embryo that were implanted or failed to be implanted in patients (n=25 pregnant and 25, non-pregnant patients). Methods: Fifty patients were accepted in the Department of Reproductive Biology of a Hospital in México City, based on the Institutional inclusion criteria for in vitro fertilization. On day 3 of development, embryos were transferred to women, and the culture medium was collected from implanted embryos (n=25, pregnant patients) and non-implanted embryos (n=25, non-pregnant). In the culture medium, RNA was isolated using TRIzol reagent. MiRNA expression was detected through RT-PCR with specific primers. Expression bands were quantified using an optic density.Results: The expression profiles were compared between pregnant and non-pregnant patients revealing a significant 5.2-fold greater expression of hsa-miR-191-5p in the former group (p ≤0.001) and a significantly higher expression of hsa-miR-24-1-5p (p =0.043) in the latter. No significant difference was found between the two groups in regard hsa-miR-21-3p or hsa-miR-372-5p (p =0.41). Conclusions: According to the results, has-miR-191-5p could possibly be a possible biomarker of adequate human embryo development. This miRNA modulated IGF2BP-1 and IGF2R, which are associated with the implantation window. On the other hand, hsa-miR-24-1-5p may be related to a poor prognosis of human embryo development.

Triploidy in a Live-Born Extremely Low Birth Weight Twin: Clinical Aspects

2020 ◽  
Author(s):  
Liliya Vakrilova ◽  
Stanislava Hitrova-Nikolova ◽  
Irena Bradinova
Keyword(s):  
Left Lung ◽  
Extremely Low Birth Weight ◽  
Clinical Aspects ◽  
Severe Respiratory Distress ◽  
Vitro Fertilization ◽  
Gestational Week ◽  
Polymerase Chain ◽  
Fluorescent Polymerase Chain Reaction ◽  
Second Twin

AbstractTriploidy is a rare chromosomal aberration characterized by a karyotype with 69 chromosomes. Triploid fetuses usually are miscarried in early pregnancy. We present a case of a triploid twin and a genetically unaffected co-twin, conceived through in vitro fertilization. A discordant growth was registered at 20 weeks of gestation. Cesarean section was performed at 355/7 gestational week. The second twin was extremely growth restricted female (780 g) with oligohydramnios and severe respiratory distress, and died at 20 hours of age. The autopsy revealed unilobar left lung, bilobar right lung, and cysts of the terminal bronchioles. Quantitative fluorescent polymerase chain reaction detected triploidy compatible pattern. So, early intrauterine growth restriction may be a sign of triploidy, which must be proven by pre or postnatal genetic testing.

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