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Spindle and chromosome configuration analysis of human biopsied versus non-biopsied embryos by confocal laser scanning microscopy following vitrification

Published online by Cambridge University Press:  07 May 2019

Katerina Chatzimeletiou Open the ORCID record for Katerina Chatzimeletiou [Opens in a new window] ,
Pierre Vanderzwalmen ,
Yannis Panagiotidis ,
Achilleas Papatheodorou ,
Alexandros Karagiannidis ,
Efstratios Kolibianakis ,
Grigoris Grimbizis ,
Nikos Prapas ,
Yannis Prapas  and
Basil C. Tarlatzis
Katerina Chatzimeletiou*
Affiliation:
Unit for Human Reproduction, 1st Department of Obstetrics & Gynaecology, Aristotle University Medical School, Papageorgiou General Hospital, Thessaloniki, Greece
Pierre Vanderzwalmen
Affiliation:
IVF Centre Prof. Zech, Bregenz, Austria, IVF Unit CHIREC, Braine l’alleud, Belgium
Yannis Panagiotidis
Affiliation:
Iakentro Advanced Medical Centre, Thessaloniki, Greece
Achilleas Papatheodorou
Affiliation:
Iakentro Advanced Medical Centre, Thessaloniki, Greece
Alexandros Karagiannidis
Affiliation:
Unit for Human Reproduction, 1st Department of Obstetrics & Gynaecology, Aristotle University Medical School, Papageorgiou General Hospital, Thessaloniki, Greece
Efstratios Kolibianakis
Affiliation:
Unit for Human Reproduction, 1st Department of Obstetrics & Gynaecology, Aristotle University Medical School, Papageorgiou General Hospital, Thessaloniki, Greece
Grigoris Grimbizis
Affiliation:
Unit for Human Reproduction, 1st Department of Obstetrics & Gynaecology, Aristotle University Medical School, Papageorgiou General Hospital, Thessaloniki, Greece
Nikos Prapas
Affiliation:
Iakentro Advanced Medical Centre, Thessaloniki, Greece
Yannis Prapas
Affiliation:
Iakentro Advanced Medical Centre, Thessaloniki, Greece
Basil C. Tarlatzis
Affiliation:
Unit for Human Reproduction, 1st Department of Obstetrics & Gynaecology, Aristotle University Medical School, Papageorgiou General Hospital, Thessaloniki, Greece
*
Address for correspondence: Katerina Chatzimeletiou. Unit for Human Reproduction, 1st Department of Obstetrics & Gynaecology, Aristotle University Medical School, Papageorgiou General Hospital, Thessaloniki, 56403, Greece. Tel. +30 231 332 3827. E-mail: katerinachatzime@hotmail.com
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Summary

The aim of this study was to investigate the effects of zona drilling and biopsy on day 3 followed by vitrification on day 5 on the cytoskeleton and development of human embryos, by analysing survival rates and spindle and chromosome configurations by fluorescence and confocal laser scanning microscopy in human biopsied and non-biopsied embryos. In total, 98 human blastocysts (50 non-biopsied and 48 following biopsy on day 3) were vitrified on day 5 using either a commercial dimethyl sulphoxide (DMSO)-free vitrification kit or increasing concentrations of DMSO/EG (5%/5–10%/10–20%/20%). Following warming, the blastocysts were allowed to recover in culture for 24 h and were immunostained with α-tubulin, acetylated tubulin, and/or γ-tubulin antibodies in combination with 4′,6-diamidino-2-phenylindole (DAPI). Labelled embryos were examined by both fluorescence and confocal laser scanning microscopy. The survival rates following warming (92% non-biopsied vs 83.3% biopsied) and the incidence of normal spindle chromosome configurations was not statistically different between the two groups (65.2% non-biopsied vs 59.2% biopsied, P>0.05). The incidence of spindle abnormalities including multipolarity, chromosome lagging, congression failure and chromosome bridging were also similar between the two groups (P>0.05). This study is the first to compare the incidence of cytoskeletal abnormalities in biopsied and non-biopsied human embryos following vitrification. We conclude that there was no significant difference in the survival rates and the incidence of spindle abnormalities between the two groups.

Keywords

Aseptic vitrificationConfocal laser scanning microscopyEmbryo biopsyHuman blastocystSpindle abnormalities
Type
Research Article
Information
Zygote , Volume 27 , Issue 3 , June 2019 , pp. 153 - 159
Copyright
© Cambridge University Press 2019 

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