Human Embryonic Behavior Observed with Time-Lapse Cinematography

Mio, Y.

doi:10.4172/2157-7420.1000143

Cited by 12 publications

(20 citation statements)

References 34 publications

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“…Dynamic analyses of TLC images demonstrated several aberrant zygotes with 1PN and uneven 2PN [7]. A previous study has reported that a third PB-like material was extruded into the perivitelline space in some zygotes with 1PN and in uneven 2PN zygotes [7]. Surprisingly, an fPN-like body formed within the third PB-like material in uneven 2PN zygotes and was reabsorbed into the cytoplasm.…”

Section: Discussionmentioning

confidence: 80%

“…A review of chromosome abnormalities in human embryos after c-IVF and ICSI identified numerous reports that used fluorescence in situ hybridization to study chromosomal status [14][15][16][17][18]. More recently, we developed an in vitro culture system for TLC to analyze human embryonic development from fertilization to the hatched blastocyst stage [7,8,[19][20][21]. Dynamic analyses of TLC images demonstrated several aberrant zygotes with 1PN and uneven 2PN [7].…”

Section: Discussionmentioning

confidence: 99%

“…More recently, we developed an in vitro culture system for TLC to analyze human embryonic development from fertilization to the hatched blastocyst stage [7,8,[19][20][21]. Dynamic analyses of TLC images demonstrated several aberrant zygotes with 1PN and uneven 2PN [7]. A previous study has reported that a third PB-like material was extruded into the perivitelline space in some zygotes with 1PN and in uneven 2PN zygotes [7].…”

Section: Discussionmentioning

confidence: 99%

“…ART procedures also routinely assess if normal fertilization has occurred by determining whether the embryo has two pronuclei (PN) and if the second polar body (2nd PB) has been extruded. Abnormally fertilized human zygotes with one pronucleus (1PN) or three pronuclei (3PN) have been identified in previous studies [1][2][3][4][5][6][7]. Several hypotheses exist to explain such aneuploidies, but the underlying causes in human ART-derived zygotes remain unknown.…”

Section: Introductionmentioning

confidence: 99%

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Diagnosis of abnormal human fertilization status based on pronuclear origin and/or centrosome number

Kai¹,

Iwata²,

Iba³

et al. 2015

J Assist Reprod Genet

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Purpose Normally fertilized zygotes generally show two pronuclei (2PN) and the extrusion of the second polar body. Conventional in vitro fertilization (c-IVF) and intracytoplasmic sperm injection (ICSI) often result in abnormal monopronuclear (1PN), tripronuclear (3PN), or other polypronuclear zygotes. In this study, we performed combined analyses of the methylation status of pronuclei (PN) and the number of centrosomes, to reveal the abnormal fertilization status in human zygotes. Method We used differences in DNA methylation status (5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC)) to discriminate between male and female PN in human zygotes. These results were also used to analyze the centrosome number to indicate how many sperm entered into the oocyte. Result Immunofluorescent analysis shows that all of the normal 2PN zygotes had one 5mC/5hmC double-positive PN and one 5mC-positive PN, whereas a parthenogenetically activated oocyte had only 5mC staining of the PN. All of the zygotes derived from ICSI (1PN, 3PN) had two centrosomes as did all of the 2PN zygotes derived from c-IVF. Of the 1PN zygotes derived from c-IVF, more than 50 % had staining for both 5mC and 5hmC in a single PN, and one or two centrosomes, indicating fertilization by a single sperm. Meanwhile, most of 3PN zygotes derived from c-IVF had a 5mC-positive PN and two 5mC/5hmC double-positive PNs, and had four or five centrosomes, suggesting polyspermy. Conclusions We have established a reliable method to identify the PN origin based on the epigenetic status of the genome and have complemented these results by counting the centrosomes of zygotes.

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Section: Discussionmentioning

confidence: 80%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Diagnosis of abnormal human fertilization status based on pronuclear origin and/or centrosome number

Kai¹,

Iwata²,

Iba³

et al. 2015

J Assist Reprod Genet

Self Cite

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“…1) [14,15]. Using this system, we have demonstrated various novel phenomena during embryonic development in human embryos from the fertilization process to the hatched blastocyst stage without any damage to the embryos [16][17][18][19][20].…”

Section: Introductionmentioning

confidence: 99%

Observation of human embryonic behavior in vitro by high‐resolution time‐lapse cinematography

Iwata

Mio²

2016

Reprod Medicine & Biology

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Assisted reproductive technology (ART) has yielded vast amounts of information and knowledge on human embryonic development in vitro; however, still images provide limited data on dynamic changes in the developing embryos. Using our high-resolution time-lapse cinematography (hR-TLC) system, we were able to describe normal human embryonic development continuously from the fertilization process to the hatched blastocyst stage in detail. Our hR-TLC observation also showed the embryonic abnormality of a third polar body (PB)-like substance likely containing a small pronucleus being extruded and resulting in single-pronucleus (1PN) formation, while our molecular biological investigations suggested the possibility that some 1PN embryos could be diploid, carrying both maternal and paternal genomes. Furthermore, in some embryos the extruded third PB-like substance was eventually re-absorbed into the ooplasm resulting in the formation of an uneven-sized, two-PN zygote. In addition, other hR-TLC observations showed that cytokinetic failure was correlated with equal-sized, multi-nucleated blastomeres that were also observed in the embryo showing early initiation of compaction. Assessment combining our hR-TLC with molecular biological techniques enables a better understanding of embryonic development and potential improvements in ART outcomes.

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Morphokinetic features in human embryos: Analysis by our original high‐resolution time‐lapse cinematography—Summary of the past two decades

Mio,

Yumoto,

Sugishima

et al. 2024

Reprod Medicine & Biology

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BackgroundThe pioneering work by Dr. Payne et al. in time‐lapse cinematography for observation of the morphokinetic features of human embryos inspired us to develop a new in vitro culture system with high‐resolution time‐lapse cinematography (hR‐TLC) back in 2001.MethodsThis in vitro culture system was capable of maintaining stable culture and was constructed on an inverted microscope stage. Embryos were observed and photographed noninvasively for an extended period, up to 7 days. The obtained images were displayed at a speed of 30 frames per second and individually analyzed.ResultsUsing hR‐TLC, human fertilization and subsequent embryonic development were visualized, revealing the time course of phenomena and many unusual dynamics.ConclusionIn this review, we summarize the results of our hR‐TLC analysis of early human embryonic development over the past 20 years. In the near future, it is expected that the vast amount of information obtained by hR‐TLC will be integrated into the AI system for further analysis and to provide feedback that will have the potential to improve clinical practice. In the era of SDGs and environmental awareness, we should be cautious about the direction in which AI can be utilized to avoid any further harm to the planet.

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Human Embryonic Behavior Observed with Time-Lapse Cinematography

Cited by 12 publications

References 34 publications

Diagnosis of abnormal human fertilization status based on pronuclear origin and/or centrosome number

Diagnosis of abnormal human fertilization status based on pronuclear origin and/or centrosome number

Observation of human embryonic behavior in vitro by high‐resolution time‐lapse cinematography

Morphokinetic features in human embryos: Analysis by our original high‐resolution time‐lapse cinematography—Summary of the past two decades

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