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Comparison of 2, 5, and 20 % O2 on the development of post-thaw human embryos

  • Embryo Biology
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

The objective of this study is to investigate the effect of 2, 5, and 20 % O2 on post-thaw day 3 human embryo culture until blastocyst stage.

Methods

One hundred fifty-five day 3 human embryos were used. One hundred twenty out of 155 embryos were recovered after thawing. Surviving embryos were distributed into 2, 5, or 20 % O2 groups and cultured for 2.5 days. At the end of culture, blastocyst formation was assessed, and then, embryos were collected for RT-qPCR or immunofluorescence analysis.

Results

Using visible blastocoel to define blastocyst formation, 58.7 % (27/46) of surviving day 3 embryos formed blastocyst at 2 % O2, 63.6 % (28/44) at 5 % O2, and 66.7 % (20/30) at 20 % O2. The difference in blastocyst formation rates was not significant. Average blastocyst cell number was 119.44 ± 11.64 at 2 % O2, 142.55 ± 22.47 at 5 % O2, and 97.29 ± 14.87 at 20 % O2. Average apoptotic rate was 4.7 % ± 0.4 % for blastocyst formed at 2 % O2, 3.5 % ± 0.7 % at 5 % O2, and 5.8 % ± 1.1 % at 20 % O2. Apoptosis rate was significantly lower for blastocysts formed at 5 % O2 (p < 0.05). Compared with gene expression levels at 5 % O2, which were arbitrarily set as “1,” 20 % O2 is associated with significantly higher expression of BAX (2.14 ± 0.47), G6PD (2.92 ± 1.06), MnSOD (2.87 ± 0.88), and HSP70.1 (8.68 ± 4.19). For all genes tested, no significant differences were found between 2 and 5 % O2.

Conclusion

The result suggests that development of cryopreserved human embryos from day 3 to blastocyst stage benefits from culture at 5 % O2.

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Acknowledgments

Thanks to Yuliang Liu (BS) and Yanhong Zeng (BS) for the technical support, Baomin Lu (BS) for the preparation of culture medium, Dr Tao Li for the advice on experimental design, and performance, and to Drs Husam Abu-Soud, Maik Huttemann, Mark Kibschull, and Michael Diamond for the analysis and comments on the manuscript as well as Rappolee lab members Drs Quanwen Li and Mohammed Abdulhasan.

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Author notes

  1. Mingmei Lin

    Present address: Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, 100191, China

Authors and Affiliations

  1. Center for Reproductive Medicine, First Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510080, China

    Yu Yang, Yanwen Xu, Chenhui Ding, Rabea Youcef khoudja, Mingmei Lin & Canquan Zhou

  2. Ob/Gyn, Wayne State University Medical School, 275 East Hancock Street, Detroit, MI, 48201, USA

    Yu Yang, Awoniyi O. Awonuga, Jing Dai, Elizabeth E. Puscheck & Daniel A. Rappolee

  3. Reproductive Sciences/Physiology, Wayne State University Medical School, 540 East Canfield, Detroit, MI, 48201, USA

    Yu Yang & Daniel A. Rappolee

  4. Key Laboratory of Reproductive Medicine of Guangdong Province, Guangzhou, China

    Yanwen Xu, Chenhui Ding, Rabea Youcef khoudja, Mingmei Lin & Canquan Zhou

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  1. Yu Yang
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  2. Yanwen Xu
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Corresponding author

Correspondence to Canquan Zhou.

Ethics declarations

Written informed consent was obtained from all participants before the usage of their embryos. The institutional review board (IRB) of the First Affiliated Hospital of Sun Yat-Sen University approved the study.

Conflict of interest

The authors declare that they have no conflict of interest.

Funding

This research was supported by grants from National Basic Research Program of China (No. 2012CB947604), Science and Information Technology of Guangzhou (No. 201300000097), DAR and EEP from the Office of the Vice President for Research at Wayne State University, and from the funding of the Mary Iacobelli Endowed Chair.

Additional information

Capsule The result suggests that development of cryopreserved human embryos from day 3 to blastocyst stage benefits from culture at 5 % O2.

Yu Yang and Yanwen Xu contributed equally to this work.

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Yang, Y., Xu, Y., Ding, C. et al. Comparison of 2, 5, and 20 % O2 on the development of post-thaw human embryos. J Assist Reprod Genet 33, 919–927 (2016). https://doi.org/10.1007/s10815-016-0693-5

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  • DOI: https://doi.org/10.1007/s10815-016-0693-5

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