Penetration of the Zona Pellucida of Nonliving Human Oocytes by Human Spermatozoa in Vitro**Supported by a grant from The Ford Foundation to the International Institute for the Study of Human Reproduction, Columbia University, and by Medical Scientist Training Fellowship Grant GM 02042 (to J. W. O.).

SummaryThe ultrastructural analysis of human oocytes at different maturation stages has only been descriptive. The aim of this study was to use a stereological approach to quantify the distribution of organelles in oocytes at prophase I (GV). Seven immature GV oocytes were processed for transmission electron microscopy and a classical manual stereological technique based on point-counting with an adequate stereological grid was used. The Kruskal–Wallis test and Mann–Whitney U-test with Bonferroni correction were used to compare the means of the relative volumes occupied by organelles in oocyte regions: cortex (C), subcortex (SC) and inner cytoplasm (IC). Here we first describe in GV oocytes very large vesicles of the smooth endoplasmic reticulum (SER), vesicles containing zona pellucida-like materials and coated vesicles. The most abundant organelles were the very large vesicles of the SER (6.9%), mitochondria (6.3%) and other SER vesicles (6.1%). Significant differences in organelle distribution were observed between ooplasm regions: cortical vesicles (C: 1.3% versus SC: 0.1%, IC: 0.1%, P = 0.001) and medium-sized vesicles containing zona pellucida-like materials (C: 0.2% versus SC: 0.02%, IC: 0%, P = 0.004) were mostly observed at the oocyte cortex, whereas mitochondria (C: 3.6% versus SC: 6.0%, IC: 7.2%, P = 0.005) were preferentially located in the subcortex and inner cytoplasm, and SER very large vesicles (IC: 10.1% versus C: 0.9%, SC: 1.67%, P = 0.001) in the oocyte inner cytoplasm. Further quantitative studies are needed in immature metaphase-I and mature metaphase-II oocytes, as well as analysis of correlations between ultrastructural and molecular data, to better understand human oocyte in vitro maturation.

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Zona pellucida birefringence and meiotic spindle visualisation of human oocytes are not influenced by IVM technology

Reproduction Fertility and Development ◽

10.1071/rd13001 ◽

2014 ◽

Vol 26 (3) ◽

pp. 407 ◽

Cited By ~ 12

Author(s):

Marjan Omidi ◽

Mohammad Ali Khalili ◽

Sareh Ashourzadeh ◽

Marzieh Rahimipour

Keyword(s):

Zona Pellucida ◽

Polar Body ◽

Refractile Body ◽

Meiotic Spindle ◽

Human Oocytes ◽

Non Invasive ◽

Safe Technology ◽

Maturation Rate

The aim of the present study was to investigate the relationship between the presence of the meiotic spindle and zona pellucida (ZP) birefringence with morphology of in vivo- and in vitro-matured human oocytes. Germinal vesicles (n = 47) and MI (n = 38) oocytes obtained from stimulated ovaries of patients undergoing intracytoplasmic sperm injection (ICSI) underwent IVM. Using a PolScope (OCTAX PolarAID; Octax, Herbon, Germany), the presence of spindles and ZP birefringence was assessed in both in vivo-matured (n = 56) and IVM (n = 56) oocytes. In addition, the morphology of each matured oocyte was evaluated microscopically. There were insignificant differences for ZP birefringence and meiotic spindle between the in vivo-matured and IVM MII oocytes. Subanalysis revealed that the rates of morphologically abnormal oocytes did not differ significantly between the two groups, except in the case of irregular shape (P = 0.001), refractile body (P = 0.001) and fragmented polar body (P = 0.03), which were higher in IVM oocytes. In the case of in vivo-matured oocytes, a significantly higher percentage of oocytes with intracytoplasmic and both intra- and extracytoplasmic abnormalities have a low birefringent ZP (P = 0.007 and P = 0.02, respectively). There was no relationship between morphological abnormalities and spindle detection. The findings suggest that clinical IVM is a safe technology that maintains the high maturation rate and integrity of oocytes. In addition, the use of the non-invasive PolScope is recommended for the detection of oocytes most suitable for ICSI.

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Superhydrophobic Soot Coated Quartz Crystal Microbalances: A Novel Platform for Human Spermatozoa Quality Assessment

Sensors ◽

10.3390/s19010123 ◽

2019 ◽

Vol 19 (1) ◽

pp. 123 ◽

Cited By ~ 12

Author(s):

Karekin Esmeryan ◽

Rumiana Ganeva ◽

Georgi Stamenov ◽

Todor Chaushev

Keyword(s):

Quality Assessment ◽

Quartz Crystal ◽

Semen Quality ◽

Human Spermatozoa ◽

Human Reproduction ◽

Quality Analysis ◽

Computer Assisted ◽

Liquid Environment ◽

Male Gametes

The functionality of human spermatozoa is a key factor for the success rate of natural human reproduction, but unfortunately the infertility progressively increases due to multifarious environmental factors. Such disquieting statistics requires the employment of sophisticated computer-assisted methods for semen quality analysis, whose precision, however, is unreliable in cases of patients with low sperm concentrations. In this study, we report a novel quartz crystal microbalance (QCM) based biosensor for in-situ quality assessment of male gametes, comprising a superhydrophobic soot coating as an interface sensing material. The soot deposition on the surface of a 5 MHz QCM eliminates the noise that normally arises upon immersion of the uncoated sensor in the test liquid environment, allowing the detection of human spermatozoa down to 1000–100,000 units/mL (1–100 ppb). Furthermore, the soot coated QCM delimitates in a highly repeatable way the immotile and motile sperm cells by inducing fundamentally distinct responses in respect to sensor sensitivity and signal trends. The obtained results reveal the strong potential of the superhydrophobic QCM for future inclusion in diverse laboratory analyses closely related to the in vitro fertilization procedures, with a final aim of gaining practical approaches for diagnoses and selection of male gametes.

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Potential health hazards of assisted human reproduction: Effects of manipulations in vitro of human oocytes and embryos on the birthweight of the resultant babies

Human Reproduction ◽

10.1093/humrep/10.6.1322 ◽

1995 ◽

Vol 10 (6) ◽

pp. 1322-1324 ◽

Cited By ~ 2

Author(s):

J. J. Tarin ◽

A. Cano

Keyword(s):

Health Hazards ◽

Human Reproduction ◽

Potential Health ◽

Human Oocytes ◽

Assisted Human Reproduction

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Effect of oocyte quality and sperm characteristics on the number of spermatozoa bound to the zona pellucida of human oocytes inseminated in vitro

Journal of In Vitro Fertilization and Embryo Transfer ◽

10.1007/bf01533760 ◽

1987 ◽

Vol 4 (4) ◽

pp. 223-227 ◽

Cited By ~ 19

Author(s):

Maha M. Mahadevan ◽

Alan O. Trounson ◽

Carl Wood ◽

John F. Leeton

Keyword(s):

Zona Pellucida ◽

Oocyte Quality ◽

Human Oocytes ◽

Sperm Characteristics

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Effects of Vitrification on Outcomes of In VivoMature, In Vitro-Mature and Immature Human Oocytes

Cellular Physiology and Biochemistry ◽

10.1159/000445564 ◽

2016 ◽

Vol 38 (5) ◽

pp. 2053-2062 ◽

Cited By ~ 9

Author(s):

Wen-yan Song ◽

Zhao-feng Peng ◽

Xue-mei Chen ◽

Hai-xia Jin ◽

Gui-dong Yao ◽

...

Keyword(s):

Zona Pellucida ◽

Occurrence Rate ◽

Aneuploidy Screening ◽

Immature Oocytes ◽

Human Oocytes ◽

Group A ◽

Mature Group ◽

Group B

Background/Aims: To observe the effects of vitrification on spindle, zona pellucida, embryonic aneuploidy and DNA injury in in vivo-maruted, in vitro-mature and immature human oocytes. Methods: Between January 2009 and February 2015, 223 immature oocytes from 450 infertile patients, and 31 in vivo-mature oocytes from 3 infertile couples were collected. Of the 223 immature oocytes, 113 were used for in vitro culture before vitrification. Some oocytes were randomly divided into in vivo-mature group (group A, n = 15), in vitro-mature group (group B, n = 88) and immature group (group C, n = 85), and then the oocytes with spindle in these three groups after freezing-thawing were selected to use for Polscope imaging, embryonic aneuploidy screening and embryo development evaluation. Other oocytes were randomly divided into group A (n = 16), group B (n = 25) and group C (n = 25) for detecting DNA injury. Results: After thawing, spindle occurrence rate, spindle Retardance value, and cleavage rate were significantly higher in groups A and B than in group C (all P < 0.05), but there were no statistical differences in fertility rate, high-quality embryo rate, blastulation rate and aneuploidy rate amongst the three groups (all P > 0.05). Zona pellucida density (ZPD) was significantly lower in group A than in groups B and C both before and after vitrification (all P < 0.05). ZPD was significantly higher after thawing than before vitrification (all P < 0.05), but zona pellucida thickness (ZPT) was not significantly changed in all the three groups (all P > 0.05). Rate of comet cells was significantly lower in group A than in groups B and C (all P < 0.01). Comet tail was significantly longer in group C than in groups B and A (all P < 0.05). Conclusion: In vivo- and in vitro-mature human oocytes are more suitable to vitrification than immature human oocytes. Spindle Retardance value has more predictive value for embryonic development potential than ZPD and ZPT.

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