[In vitro parthenogenesis in the human species].
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Parthenogenesis
Cleavage (geology)
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Although successful embryo development is dependent upon genetic and epigenetic contributions from both the male and female, the male potential to adversely affect embryo development has been scarcely studied. It is unclear whether the sperm variation among different males would affect the outcome of oocyte evaluation by embryo development following fertilization. In the present study, variation in the developmental potential of mouse embryos was first compared between in vitro fertilization with epididymal spermatozoa from different males and Sr(2+) parthenogenetic activation using oocytes of different qualities, and then the effect of male on fertilization and embryo development was examined using randomly chosen oocytes and spermatozoa from cauda epididymidis, vas deferens or electro-ejaculates. Rates of fertilization and blastocyst formation were significantly higher with spermatozoa from cauda epididymidis or vas deferens than with ejaculated spermatozoa. Rates of embryonic development differed significantly between different males, but not between different ejaculates of the same male. Analysis of standard errors of means and coefficients of variance indicated that as long as multiple males were involved, the variation in oocyte fertilization/activation and blastocyst formation was always higher after fertilization than after Sr(2+) parthenogenetic activation whether spermatozoa were collected from epididymidis, vas deferens or ejaculates and regardless of oocyte qualities. It is concluded that (1) epididymal mouse spermatozoa fertilize more oocytes than ejaculated spermatozoa under identical experimental conditions; (2) like farm animals, the mice also show a remarkable male effect on the developmental potential of in vitro produced embryos although they are supposed to be less genetically diverse; (3) parthenogenetic activation is recommended for assessment of oocyte quality to exclude the effect of male.
Parthenogenesis
Vas deferens
Oocyte activation
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To investigate the effect of the membranes of mouse oocyte on fertilization in vitro as a model using oocyte of mouse after vitrification.Under room temperature(25±0.5)℃,oocytes were vitrified in OPS by two steps vitification method with cryprotectant solutions-that is,10%EG+10%DMSO and EDFS30.Oocytes were firstly pretreated in 10% EG+10% DMSO for 30 s,then exposed to EDFS30 for 25 s.The results showed that the cleavage rate of vitrificated oocytes were significantly lower(46.67% vs.86.06%)(P 0.05) than that of controls.The average number of sperm combined on the surface of the oocytes(10.70vs.10.81) and the average number of pronuclears in oocytes(2.49 vs.2.59) were all similar(P 0.05) between vitrificated and no-treated oocytes.After partial zona pellucida incision by piezo micromanipulation(ZIP),the cleavage rate of the vitrificated oocytes was similar(84.73% vs.91.19%)(P 0.05) with that of controls.In conclusion,the change of the zona pellucida of vitrificated mouse oocytes was one of the most important reasons to reduce the fetilization rate.
Cleavage (geology)
Vitrification
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Abstract Regardless of whether fertilization occurs in vivo or in vitro, polyspermic penetration of the human oocyte is a not infrequent cause of reproductive failure. This report describes the occurrence of human eggs fertilized by two spermatozoa and the variable developmental potentials expressed by the resultant embryos. The cellular and subcellular events that characterize the pronuclear and early cleavage stages of preimplantation embryogenesis are discussed with respect to the ability of such embryos to progress to implantation.
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Effects of different time ultraviolet(UV) irradiation combined with fluorescence Hoechst33342 on the cleavage and blastocyst development of parthenogenic and in vitro fertilization embryos in bovine were investigated. The results showed that the cleavage and blastocyst development of parthenogenic embryos derived from oocytes irradiated with UV 30s and 40s were significantly lower than those of oocytes irradiated with UV 0 s, 10 s and 20,s ( P 0.01). It indicated that irradiating oocytes with UV longer than 20s decreased the developmental potential of parthenogenic embryos significantly.In in vitro fertilization research, the cleavage rate of embryos derived from the irradiated oocytes suffered from UV irradiation 30 s and 40 s are significantly lower than that suffered from UV irradiation 10 s and 20 s ( P 0.05),furthermore,the blastocyst development rate had significant difference between 20s and 10s.The results indicated that irradiating oocytes with UV no longer than 20s could obtain the cleavage and blastocyst developmental potential of bovine embryos from parthenogenic and in vitro fertilization.
Parthenogenesis
Cleavage (geology)
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[Objective] The aim of the study was to seek optimum conditions for in vitro embryo production.[Method]With cow oocyte as experiment material,the effects of different temperature and transport time on the mature cow oocyte were studied;and the effects of parthenogenesis active and in vitro fertilization on cow oocyte embryonic development in the same condition were compared.[Result] The study showed that the mature rate of the oocyte after transport in 37-42 ℃(11.3%)was obviously lower than those in 20-30 ℃ and 30-37 ℃(79.3%,85.1%),the mature rate of oocyte after 6 h transport(53.2%) was much lower than that after 4h and 4-6 h(85.6%,77.8%),which were no obvious difference between the latter two.Under the same condition,parthenogenesis active and in vitro fertilization cleavage rate(72.41%,68.18%),8 Cell rate(45.24%,47.22%),blastocyte rate(25.60%,18.89%) was no obvious difference.[Conclusion]The storage of ovaries back to the laboratory within 4 h at about 30 ℃ was optimum.Under the same conditions,the development rate of parthenogenesis active and in vitro fertilization had no obvious difference.
Parthenogenesis
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A total of 518 normal-appearing, meiotically mature human oocytes that were judged unfertilized after insemination in vitro were examined for sperm penetration by conventional fluorescence and laser scanning confocal microscopy with DNA-specific probes. A similar analysis was performed on 29 single pronuclear oocytes that were presumed to originate by spontaneous (parthenogenetic) activation. The results demonstrate that 22% of the unfertilized oocytes and 52% of the presumed parthenogenetic oocytes were actually penetrated. Sperm penetration occurred in both normozoospermic and male factor cases. The findings indicate the importance of penetration analysis in determining the causes of fertilization failure that may reside with the male or female gamete, especially when assessing the utility of and necessity for assisted fertilization in subsequent attempts. The results also suggest that the cytoplasmic capacity to decondense sperm DNA may decline more rapidly than the ability of the oocyte to be penetrated and to mount an effective block to polyspermy.
Polyspermy
Gamete
Parthenogenesis
Pronucleus
Capacitation
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Abstract Success rates of superovulation in response to gonadotropic hormone treatment and in vitro fertilization (ie, mitotic cleavage following insemination) of mouse eggs from outbred CD‐1, hybrid CB6F l , or hybrid B6CBAF 1 , mice were compared using either a mouse inseminationmedium, modified Krebs‐Ringer‐bicarbonate (m‐KRB), or a human insemination medium, Ham's F10 nutrient mixture. Inseminations were performed in either organ culture dishes or screw‐top, flat‐side tissue culture tubes. Mean superovulation rates (± SD) were 24.2 (5.1) for CD‐1, 33.0 (5.8) for CB6F 1 , and 16.3 (6.6) for B6CBAF 1 mice. For in vitro cleavage the best combination of mouse strain, insemination medium, and culture container was achieved using CB6F 1 , mice, m‐KRB medium, and culture tubes. However, Ham's medium used with either hybrid mouse strain was shown to be employable for fertilization of mouse eggs in vitro as a quality control assay and/or experimental model system for testing the human in vitro fertilization procedure.
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The effect of oocyte quality on the pronuclear (PN) formation and subsequent development of embryos in local cattle was studied. Cattle oocytes matured and fertilized in vitro were unable to acquire developmental competence unless matured with intact cumulus cells. The presence of cumulus cells promoted normal fertilization with proper pronuclear (2PN) formation. In the present study, the characteristics and quality of cumulus-oocyte-complexes influenced the pronuclear formation but not subsequent cleavage and blastocyst formation. The oocytes from Grade A, Grade B and Grade B' were capable of fertilization and development under in vitro conditions. Fertilization rates were significantly different (p 2PN) was 18.8% in Grade B' oocytes but none in Grades A and B oocytes. The mean percentages of cleavage and blastocyst rates were not significant (p > 0.05) in Grades A, B and B' (71.6, 74.9 and 73.6%, respectively, and 10.5, 10.4 and 7.4%, respectively). In conclusion, this study indicated that the compactness of cumulus cells surrounding the oocytes influenced the pronuclear formation of cattle oocytes but not cleavage and blastocyst rates.
Keywords: pronuclear formation; blastocyst; cumulus-oocyte complex; in vitro fertilization; cleavage; cattle embryo
Cleavage (geology)
Embryo quality
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Capacitation
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HEPES
Parthenogenesis
Cleavage (geology)
Bicarbonate
Human chorionic gonadotropin
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