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Cryopreservation of incomplete compacted morulae and preliminary biopsy of excluded fragments

Published online by Cambridge University Press:  15 August 2019

Maryna Petrushko ,
Taisiia Yurchuk Open the ORCID record for Taisiia Yurchuk [Opens in a new window] ,
Volodymyr Piniaiev  and
Natalia Buderatska
Maryna Petrushko
Affiliation:
Institute for Problems of Cryobiology and Cryomedicine of the NAS of Ukraine, Kharkiv, Ukraine ART-clinic for Reproductive Medicine, Kharkiv, Ukraine
Taisiia Yurchuk*
Affiliation:
Institute for Problems of Cryobiology and Cryomedicine of the NAS of Ukraine, Kharkiv, Ukraine
Volodymyr Piniaiev
Affiliation:
Institute for Problems of Cryobiology and Cryomedicine of the NAS of Ukraine, Kharkiv, Ukraine ART-clinic for Reproductive Medicine, Kharkiv, Ukraine
Natalia Buderatska
Affiliation:
Institute for Problems of Cryobiology and Cryomedicine of the NAS of Ukraine, Kharkiv, Ukraine IGR Medical Centre, Kiev, Ukraine
*
Address for correspondence: Taisiia Yurchuk, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, 23 Pereyaslavska str., Kharkiv 61015, Ukraine. Tel: +380 57 373 4143. Fax: +380 57 373 3084. E-mail: taisiya.yur@gmail.com
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Summary

The complexity of predicting embryo development potential at the cleavage stages and the emergence of epigenetic risks during prolonged in vitro culture of pre-implantation embryos made it more advantageous to transfer embryos at the morula stage to the uterine cavity. The criteria for estimating embryos at this stage that allow prediction of cryopreservation outcomes have been poorly described. All day 4 embryos (n = 224) were graded 1, 2, 3, 4 or 5 according to blastomere compaction degree (BCD = 100, 75, 50, 25 or 0%, respectively) and the survival and blastocyst formation rate of these morulae were studied after cryopreservation. An inverse dependence was found between survival rate and BCD. Excluded fragments were characterized by low osmotic reaction during exposure to cryoprotective medium and, after freeze-thawing, they were destroyed. As damaged necrotic areas of the embryo can affect their further development rate we proposed blastomeres and biopsy fragments of incomplete compacted morula be removed before embryo cryopreservation. This step led to significant increase in the post-thawing survival rate up to 93.1 ± 4.1%, 75 ± 8.8% and blastocyst formation rate up to 85.2 ± 10.4%, 59.4 ± 5.2% in grade 2 and grade 3 embryos, respectively. There was no significant difference in grade 4 embryos. Therefore the removal of blastomeres and biopsy fragments in incomplete compacted morulae can improve cryopreservation outcomes of grade 2 and grade 3 embryos with BCD.

Keywords

BiopsyBlastomere compaction degreeCryopreservationIncompleteMorula
Type
Research Article
Information
Zygote , Volume 27 , Issue 6 , December 2019 , pp. 386 - 391
Copyright
© Cambridge University Press 2019 

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