Assisted reproductive research: Laser assisted hatching and spindle detection (spindle view technique)

2004 ◽  
Vol 52 (1) ◽  
pp. 113-123 ◽  
Author(s):  
Katalin Kanyó ◽  
J. Konc ◽  
L. Solti ◽  
S. Cseh
Keyword(s):  
Animal Studies ◽  
Harmful Effect ◽  
Animal Experiments ◽  
Oocyte Quality ◽  
Assisted Hatching ◽  
Safe Procedure ◽  
Living Mouse ◽  
Reproductive Techniques ◽  
Developmental Capacity

Animal experiments are very important for the development of new assisted reproductive techniques (ART) for use in human and animal reproductive medicine. Most technical aspects of reproductive manipulation of humans and animals are very similar, and many components of successful human ART used nowadays have been derived from animal studies. In this study we examined (1) the use of 'non-contact' laser for assisted hatching, (2) whether spindles in living mouse oocytes could safely be imaged/examined by polarisation microscope (polscope) and (3) the influence of environment (e.g. temperature, in vitro culture, etc.) on spindle detection/visualisation. The data of the study presented here show that (1) laser assisted hatching (AH) is a fast, very accurate and safe procedure without any harmful effect on embryo development and it can support very effectively the implantation of embryos, (2) the use of polscope facilitates the evaluation of oocyte quality and the selection of oocytes with spindle, (3) by monitoring the spindle position during intracytoplasmic sperm injection (ICSI), we can reduce spindle damage and increase the chance of fertilisation. Further studies are underway to test the hypothesised connection between spindle birefringence and developmental capacity of oocytes/embryos.

scholarly journals Comparing four laboratory three-parent techniques to construct human aged non-surrounded nucleolus germinal vesicle oocytes: A case-control study

2020 ◽  
Author(s):  
Sara Darbandi ◽  
Mahsa Darbandi ◽  
Ashok Agarwal ◽  
Hamid Reza Khorram khorshid ◽  
Mohammad Reza Sadeghi ◽  
...  
Keyword(s):  
Case Control Study ◽  
Germinal Vesicle ◽  
Oocyte Donation ◽  
Case Control ◽  
Assisted Reproductive Technique ◽  
Oocyte Competence ◽  
Reproductive Techniques ◽  
Developmental Capacity ◽  
Control Study

Background: The three-parent assisted reproductive technique may increase oocyte competence. Objective: In this case-control study, the suitability of germinal vesicle transfer (GVT), synchronous ooplasmic transfer (sOT), asynchronous ooplasmic transfer using cryopreserved MII oocyte (caOT), and asynchronous ooplasmic transfer using waste MII oocyte (waOT) for maturation of the human-aged non-surrounded nucleolus germinal vesicle-stage (NSN-GV) oocyte were investigated. Materials and Methods: NSN-GV oocytes were subjected to four methods: group A (GVT), B (sOT), C (caOT) D (waOT), and E (Control). The fusion rates, MI, MII, ICSI observations and cleavage at 2-cell, 4-cell, and 8-cell stages were compared in the groups. Results: In GVT, none of the oocytes fused. In sOT, all oocytes fused, 20 achieved the MI, 14 progressed to MII, 8 fertilized, 6 cleaved and 5, 4, and 3 achieved the 2- cells, 4-cells and 8-cells, respectively. In caOT, all oocytes fused and achieved the MI, 8 progressed to MII and fertilized, 6 cleaved and 6, 5, and 5 achieved the 2-cells, 4- cells, and 8-cells respectively. In waOT, all oocytes fused, 5 and 3 progressed to MI and MII, respectively, but only one fertilized, cleaved and reached a 4-cells stage. In group E, 6 and 2 oocytes progressed to MI and MII, respectively, and only one fertilized but arrested at the zygote stage. caOT had the highest survival rate when compared to sOT (p = 0.04), waOT (p = 0.002), and control (p = 0.001). Conclusion: The caOT method was beneficial over sOT, waOT, and GVT in supplementing the developmental capacity of human-aged NSN-GV oocytes. Key words: Assisted reproductive techniques, In vitro oocyte maturation techniques, Nuclear transfer techniques, Oocytes, Oocyte donation.

scholarly journals Oocyte maturation and quality: role of cyclic nucleotides

Reproduction ◽  
2016 ◽  
Vol 152 (5) ◽  
pp. R143-R157 ◽  
Author(s):  
R B Gilchrist ◽  
A M Luciano ◽  
D Richani ◽  
H T Zeng ◽  
X Wang ◽  
...  
Keyword(s):  
Cyclic Nucleotides ◽  
Treatment Options ◽  
Oocyte Quality ◽  
Fertility Treatment ◽  
Lower Efficiency ◽  
Oocyte Meiosis ◽  
Developmental Capacity ◽  
Oocyte Meiotic Maturation

The cyclic nucleotides, cAMP and cGMP, are the key molecules controlling mammalian oocyte meiosis. Their roles in oocyte biology have been at the forefront of oocyte research for decades, and many of the long-standing controversies in relation to the regulation of oocyte meiotic maturation are now resolved. It is now clear that the follicle prevents meiotic resumption through the actions of natriuretic peptides and cGMP – inhibiting the hydrolysis of intra-oocyte cAMP – and that the pre-ovulatory gonadotrophin surge reverses these processes. The gonadotrophin surge also leads to a transient spike in cAMP in the somatic compartment of the follicle. Research over the past two decades has conclusively demonstrated that this surge in cAMP is important for the subsequent developmental capacity of the oocyte. This is important, as oocyte in vitro maturation (IVM) systems practised clinically do not recapitulate this cAMP surge in vitro, possibly accounting for the lower efficiency of IVM compared with clinical IVF. This review particularly focuses on this latter aspect – the role of cAMP/cGMP in the regulation of oocyte quality. We conclude that clinical practice of IVM should reflect this new understanding of the role of cyclic nucleotides, thereby creating a new generation of ART and fertility treatment options.

scholarly journals Proteomes of Animal Oocytes: What Can We Learn for Human Oocytes in theIn VitroFertilization Programme?

2014 ◽  
Vol 2014 ◽  
pp. 1-11 ◽  
Author(s):  
Irma Virant-Klun ◽  
Jeroen Krijgsveld
Keyword(s):  
Assisted Reproduction ◽  
Animal Studies ◽  
Proteome Analysis ◽  
Oocyte Quality ◽  
Oocyte Development ◽  
Human Oocytes ◽  
Mammalian Oocyte

Oocytes are crucial cells for mammalian reproduction, yet the molecular principles underlying oocyte development are only partially understood. Therefore, contemporary proteomic approaches have been used increasingly to provide new insights into oocyte quality and maturation in various species such as mouse, pig, and cow. Especially, animal studies have helped in elucidating the molecular status of oocytes duringin vitromaturation and other procedures of assisted reproduction. The aim of this review is to summarize the literature on mammalian oocyte proteome and secretome research in the light of natural and assisted reproduction and on lessons to be learned for human oocytes, which have so far remained inaccessible for proteome analysis.

scholarly journals Pyometra does not affect some molecular quality-related parameters of canine oocytes

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Piotr Pawlak ◽  
Ewelina Warzych ◽  
Zofia E. Madeja ◽  
Tomasz Nowak ◽  
Dorota Lechniak
Keyword(s):  
Mrna Level ◽  
Oocyte Quality ◽  
Potential Effect ◽  
Animal Cloning ◽  
Maternal Effect Genes ◽  
Negative Effect ◽  
Reproductive Techniques ◽  
Two Parameters

AbstractCystic endometrial hyperplasia-pyometra complex (CEH/P) significantly perturbs the reproductive performance of affected bitches and ovariohysterectomy (OHE) is a commonly applied treatment. Thus the only way to take advantage of the genetic potential of valuable females is application of assisted reproductive techniques (ART) mainly in vitro embryo production (IVP) or in some exceptional cases animal cloning by somatic cell nuclear transfer (SCNT). The aim of our study was to examine a potential effect of the CEH/P status on the quality of oocytes from females subjected to OHE. In total, 828 immature oocytes collected from ovaries of 33 bitches (21 control, 12 CEH/P) were subjected to genetic analyses (mRNA expression of two maternal-effect genes: GDF-9, OCT4 and mitochondrial DNA – mtDNA - content). Oocytes of CEH/P females were characterized by a higher mtDNA content (471 696) than gametes of their healthy counterparts (368 175; p<0,005). Transcripts for the two genes were detected in all samples and the mRNA level was not affected by the CEH/P status. In conclusion, the CEH/P complex does not exert a negative effect on oocyte quality reflected by the two parameters examined in this study.

Mechanistic insights into the reduced developmental capacity of in vitro matured oocytes and importance of cumulus cells in oocyte quality determination

2020 ◽  
Vol 235 (12) ◽  
pp. 9743-9751
Author(s):  
Ang Li ◽  
Feng Wang ◽  
Li Li ◽  
Li‐Hua Fan ◽  
Tie‐Gang Meng ◽  
...  
Keyword(s):  
Cumulus Cells ◽  
Oocyte Quality ◽  
Developmental Capacity

scholarly journals Management of ovarian functions by melatonin

2021 ◽  
Vol 2 ◽  
pp. 16
Author(s):  
Chandana Haldar ◽  
Jayita Pal Chowdhury
Keyword(s):  
Defense Mechanism ◽  
De Novo ◽  
Oocyte Quality ◽  
Melatonin Receptors ◽  
Pars Tuberalis ◽  
Reproductive Techniques ◽  
Molecular Machinery ◽  
Lh Release ◽  
Ovulatory Follicles

Extensive research has unraveled a niche for melatonin that is of great significance for the female reproductive physiology. The potency of melatonin as an antioxidant, anti-inflammatory, and anti-apoptotic agent is being utilized to benefit female reproductive anomalies. Melatonin receptors have been localized in the Supra Chaismatic Nucleus (SCN), pars tuberalis (PT), and the gonads suggesting the regulation of reproduction by melatonin not only at a higher level but also on the gonads through complex interrelated mechanisms. Melatonin secreted by the pineal gland acts on the hypothalamus to regulate gonadotropin-releasing hormone and subsequently gonadotropin (FSH/LH) release from the PT. However, the de novo synthesis of this indoleamine reported in the gonads gave rise to the idea of a more localized action. The mammalian ovary has all the molecular machinery required for the biosynthesis of melatonin and interestingly concentration of melatonin in the follicular fluid of pre-ovulatory follicles is much higher than circulatory melatonin even in humans. This locally produced melatonin has been shown to modulate various pathways governing ovarian steroidogenesis. Further, melatonin and its receptors play a significant role in antioxidant defense mechanism of ovary for follicular growth and maturation. Exposure to stress strongly influences hypothalamic-pituitary-adrenal axis and elevated glucocorticoid levels suppress various ovarian functions including implantation thereby pregnancy. Melatonin acts antagonistically with glucocorticoids, making it crucial for the management of the female reproductive functions/dysfunctions. Usage of melatonin during in vitro fertilization (IVF) procedures has been found to improve oocyte quality, survival, and fecundity. Therefore, in future, melatonin can be implicated as preferable therapeutic especially in IVF and assisted reproductive techniques.

scholarly journals Effects of Exogenous Androgens on Platelet Activity and Their Thrombogenic Potential in Supraphysiological Administration: A Literature Review

2021 ◽  
Vol 10 (1) ◽  
pp. 147
Author(s):  
Adrian Eugen Roşca ◽  
Ana-Maria Vlădăreanu ◽  
Alina Mititelu ◽  
Bogdan Ovidiu Popescu ◽  
Corin Badiu ◽  
...  
Keyword(s):  
Animal Studies ◽  
Case Reports ◽  
Animal Experiments ◽  
Platelet Activity ◽  
Androgen Excess ◽  
Life Threatening ◽  
Thrombogenic Potential ◽  
Androgenic Steroids

Anabolic androgenic steroids (AAS), simply called “androgens”, represent the most widespread drugs used to enhance performance and appearance in a sporting environment. High-dosage and/or long-term AAS administration has been associated frequently with significant alterations in the cardiovascular system, some of these with severe endpoints. The induction of a prothrombotic state is probably the most life-threatening consequence, suggested by numerous case reports in AAS-abusing athletes, and by a considerable number of human and animal studies assessing the influence of exogenous androgens on hemostasis. Despite over fifty years of research, data regarding the thrombogenic potential of exogenous androgens are still scarce. The main reason is the limited possibility of conducting human prospective studies. However, human observational studies conducted in athletes or patients, in vitro human studies, and animal experiments have pointed out that androgens in supraphysiological doses induce enhanced platelet activity and thrombopoiesis, leading to increased platelet aggregation. If this tendency overlaps previously existing coagulation and/or fibrinolysis dysfunctions, it may lead to a thrombotic diathesis, which could explain the multitude of thromboembolic events reported in the AAS-abusing population. The influence of androgen excess on the platelet activity and fluid–coagulant balance remains a subject of debate, urging for supplementary studies in order to clarify the effects on hemostasis, and to provide new compelling evidence for their claimed thrombogenic potential.

scholarly journals Human adipose-derived stromal cells transplantation prolongs reproductive lifespan on mouse models of mild and severe premature ovarian insufficiency

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Giulia Salvatore ◽  
Massimo De Felici ◽  
Susanna Dolci ◽  
Cosimo Tudisco ◽  
Rosella Cicconi ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Human Adipose Tissue ◽  
Oocyte Quality ◽  
Premature Ovarian Insufficiency ◽  
Ovarian Insufficiency ◽  
Oocyte Competence ◽  
Adipose Derived Stromal Cells ◽  
Developmental Capacity ◽  
And Performance

Abstract Background Although recent studies have investigated the ability of Mesenchymal Stromal Cells (MSCs) to alleviate short-term ovarian damage in animal models of chemotherapy-induced Premature Ovarian Insufficiency (POI), no data are available on reproductive lifespan recovery, especially in a severe POI condition. For this reason, we investigated the potential of MSCs isolated from human adipose tissue (hASCs), since they are easy to harvest and abundant, in ameliorating the length and performance of reproductive life in both mild and severe chemotherapy-induced murine POI models. Methods Mild and severe POI models were established by intraperitoneally administering a light (12 mg/kg busulfan + 120 mg/kg cyclophosphamide) or heavy (30 mg/kg busulfan + 120 mg/kg cyclophosphamide) dose of chemotherapy, respectively, in CD1 mice. In both cases, a week later, 1 × 106 hASCs were transplanted systemically through the tail vein. After four additional weeks, some females were sacrificed to collect ovaries for morphological evaluation. H&E staining was performed to assess stroma alteration and to count follicle numbers; immunofluorescence staining for αSMA was used to analyse vascularization. Of the remaining females, some were mated after superovulation to collect 2-cell embryos in order to evaluate their pre-implantation developmental capacity in vitro, while others were naturally mated to monitor litters and reproductive lifespan length. F1 litters’ weight, ovaries and reproductive lifespan were also analysed. Results hASC transplantation alleviated ovarian weight loss and size decrease and reduced alterations on ovarian stroma and vasculature, concurrently preventing the progressive follicle stockpile depletion caused by chemotherapy. These effects were associated with the preservation of the oocyte competence to develop into blastocyst in vitro and, more interestingly, with a significant decrease of chemotherapy-induced POI features, like shortness of reproductive lifespan, reduced number of litters and longer time to plug (the latter only presented in the severe POI model). Conclusion Human ASC transplantation was able to significantly reduce all the alterations induced by the chemotherapeutic treatment, while improving oocyte quality and prolonging reproductive functions, thus counteracting infertility. These results, strengthened by the use of an outbred model, support the potential applications of hASCs in women with POI, nowadays mainly induced by anticancer therapies. Graphic abstract

Decellularized bone extracellular matrix in skeletal tissue engineering

2020 ◽  
Vol 48 (3) ◽  
pp. 755-764
Author(s):  
Benjamin B. Rothrauff ◽  
Rocky S. Tuan
Keyword(s):  
Tissue Engineering ◽  
Bone Regeneration ◽  
Tissue Regeneration ◽  
Animal Studies ◽  
Tumor Resection ◽  
Regenerative Capacity ◽  
Skeletal Tissue ◽  
Large Bone

Bone possesses an intrinsic regenerative capacity, which can be compromised by aging, disease, trauma, and iatrogenesis (e.g. tumor resection, pharmacological). At present, autografts and allografts are the principal biological treatments available to replace large bone segments, but both entail several limitations that reduce wider use and consistent success. The use of decellularized extracellular matrices (ECM), often derived from xenogeneic sources, has been shown to favorably influence the immune response to injury and promote site-appropriate tissue regeneration. Decellularized bone ECM (dbECM), utilized in several forms — whole organ, particles, hydrogels — has shown promise in both in vitro and in vivo animal studies to promote osteogenic differentiation of stem/progenitor cells and enhance bone regeneration. However, dbECM has yet to be investigated in clinical studies, which are needed to determine the relative efficacy of this emerging biomaterial as compared with established treatments. This mini-review highlights the recent exploration of dbECM as a biomaterial for skeletal tissue engineering and considers modifications on its future use to more consistently promote bone regeneration.

Modulation of Matrix Metalloproteinases by Plant-Derived Products

2020 ◽  
Vol 20 ◽  
Author(s):  
Nur Najmi Mohamad Anuar ◽  
Nurul Iman Natasya Zulkafali ◽  
Azizah Ugusman
Keyword(s):  
Clinical Trials ◽  
Natural Products ◽  
Matrix Metalloproteinases ◽  
Animal Studies ◽  
Current Knowledge ◽  
In Vitro Models ◽  
In Vivo Studies ◽  
Extracellular Matrix Components

: Matrix metalloproteinases (MMPs) are a group of zinc-dependent metallo-endopeptidase that are responsible towards the degradation, repair and remodelling of extracellular matrix components. MMPs play an important role in maintaining a normal physiological function and preventing diseases such as cancer and cardiovascular diseases. Natural products derived from plants have been used as traditional medicine for centuries. Its active compounds, such as catechin, resveratrol and quercetin, are suggested to play an important role as MMPs inhibitors, thereby opening new insights into their applications in many fields, such as pharmaceutical, cosmetic and food industries. This review summarises the current knowledge on plant-derived natural products with MMP-modulating activities. Most of the reviewed plant-derived products exhibit an inhibitory activity on MMPs. Amongst MMPs, MMP-2 and MMP-9 are the most studied. The expression of MMPs is inhibited through respective signalling pathways, such as MAPK, NF-κB and PI3 kinase pathways, which contribute to the reduction in cancer cell behaviours, such as proliferation and migration. Most studies have employed in vitro models, but a limited number of animal studies and clinical trials have been conducted. Even though plant-derived products show promising results in modulating MMPs, more in vivo studies and clinical trials are needed to support their therapeutic applications in the future.

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