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The impacts of laser zona thinning on hatching and implantation of vitrified-warmed mouse embryos

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Abstract

Embryo vitrification has advantages in assisted reproduction yet it also induces zona hardening. Laser zona thinning (LZT) is considered as a solution yet its efficacy and security have not been well studied. In this study, we used vitrified-warmed morulae from 2-month-old and 10-month-old ICR female mice as model to investigate the impacts that LZT treatment brings to the in vitro hatching process and implantation by analyzing hatching rate, implantation rate, and blastocyst quality. The results showed that the fully hatched rate was significantly higher after LZT treatment for both young (25.7% vs. 16.2%, P < 0.05) and aged (36.6% vs. 13.2%, P < 0.01) mice. For zona-thinned morulae in young mice, its onset of hatching occurred earlier (28.6% vs. 8.8%, P < 0.01) at D4 and with a greater percentage of U-shaped hatching at D5 (48.3% vs. 33.0%, P < 0.05). LZT treatment did not induce expression change of apoptosis-related genes in all groups (P > 0.05), but for young mice, the total cell number of day 5 blastocyst in zona-thinned group was significantly less than that of the control group (40.6 ± 5.1 vs. 59.9 ± 14.5, P < 0.01). At last, there was an increasing implantation rate in zona-thinned compared to the control group for young (63.8% vs. 52.5%, P > 0.05) and aged (55.6% vs. 47.2%; P > 0.05) mice after embryos were bilaterally transferred in the same recipient. In conclusion, the significant increase of fully hatched rate after LZT treatment is related to the advanced onset of hatching as well as the enhancement of superior hatching structure, and LZT also lead to a better implantation after embryo transfer.

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Funding

This work was supported by the National Transgenic Creature Breeding Grand Project (2016ZX08008-003) and National Natural Science Foundation Project of China (No. 31101714 &31372307).

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  1. Zhengyuan Huang and Jinghao Liu contributed equally to this work.

Authors and Affiliations

  1. National Engineering Laboratory for Animal Breeding and Key Laboratory of Animal Genetics, Breeding and Reproduction, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing, China

    Zhengyuan Huang, Lei Gao, Qingrui Zhuan, Yuxi Luo, Shien Zhu & Xiangwei Fu

  2. Laboratory Animal Center, Peking University, Beijing, China

    Jinghao Liu

  3. BGI Clinical Laboratories (Shenzhen) Co., Ltd, Shenzhen, China

    Kaiyu Lei

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  1. Zhengyuan Huang
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Correspondence to Xiangwei Fu.

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The use of the animals and the experimental procedures were approved by the Animal Care Committee at China Agricultural University. The experiment was conducted at China Agricultural University. All methods were carried out in accordance with the approved guidelines.

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The authors declare that they have no conflict of interest.

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Huang, Z., Liu, J., Gao, L. et al. The impacts of laser zona thinning on hatching and implantation of vitrified-warmed mouse embryos. Lasers Med Sci 34, 939–945 (2019). https://doi.org/10.1007/s10103-018-2681-8

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  • DOI: https://doi.org/10.1007/s10103-018-2681-8

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