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Effect of culture medium volume and embryo density on early mouse embryonic development: Tracking the development of the individual embryo

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

Abstract

Purpose

To determine the optimal volume or density of embryos for the well-of-the-well (WOW) system in order to track the development of individual embryos and to determine whether the WOW system can reverse the negative impact of culturing embryos singly.

Methods

(1) Mouse embryos (groups of nine at the 2-cell stage) were cultured in 6.25 μl, 12.50 μl, 25.00 μl and 50.00 μl of droplets of culture medium under paraffin oil; (2) Groups of three, six, nine and twelve embryos at the 2-cell stage were cultured in 50 μl of droplet of culture medium under paraffin oil; (3) Groups of nine embryos at the 2-cell stage were cultured in 50 μl of droplet under paraffin oil with or without nine micro-wells made on the bottom of the Petri dish into each of which were placed one of the nine embryos (WOW system). Also single 2-cell stage embryos was cultured individually in 5.5 μl of droplet of culture medium under paraffin oil with or without a single micro-well made on the bottom of the Petri dish (WOW system for single culture). At the end of culture, the percentages of blastocyst development, hatching and hatched blastocysts were compared in each group. The blastocysts were fixed for differential staining.

Results

The blastocyst development was significantly higher (P < 0.05) when nine embryos were cultured in 50 μl of droplet of culture medium compared with other volumes. The blastocyst development was significantly reduced (P < 0.05) in single embryo culture compared to group embryo culture with or without the WOW system. The blastocyst development was not improved when single embryo cultured individually in a micro-well was compared to single embryo cultured individually without micro-well. The total cell numbers of blastocysts were significantly higher in group embryo culture than single embryo culture regardless of whether the WOW system was used. In addition, the total cell numbers of blastocysts were significantly higher (P < 0.05) in single embryo culture with the WOW system than without.

Conclusions

Group embryo culture is superior to single embryo culture for blastocyst development. The WOW system with 50 μl of droplet of culture medium can be used to track the individual development of embryo cultured in groups while preserving good embryonic development. The reduced embryonic development with single embryo culture cannot be ameliorated by the WOW system.

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Declaration of Interest

There is no conflict of interest that could affect the impartiality of the research reported.

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, McGill University, Montreal, QC, Canada

    Shan-Jun Dai, Chang-Long Xu, Jeffrey Wang & Ri-Cheng Chian

  2. The Reproductive Medical Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, People’s Republic of China

    Shan-Jun Dai & Ying-Pu Sun

  3. Royal Victoria Hospital, Women’s Pavilion F3, 687 Pine Avenue West, Montreal, QC, Canada, H3A 1A1

    Ri-Cheng Chian

Authors
  1. Shan-Jun Dai
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  2. Chang-Long Xu
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  3. Jeffrey Wang
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  4. Ying-Pu Sun
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  5. Ri-Cheng Chian
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Corresponding authors

Correspondence to Ying-Pu Sun or Ri-Cheng Chian.

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Capsule

Group embryo culture is superior to single embryo culture in terms of early embryonic development. The well-of-the-well (WOW) system with 50 μl culture medium droplets can be used to track the individual development of group cultured embryo with better embryonic development. However, the impairment of embryonic development with single embryo culture is not overcome using the WOW system.

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Dai, SJ., Xu, CL., Wang, J. et al. Effect of culture medium volume and embryo density on early mouse embryonic development: Tracking the development of the individual embryo. J Assist Reprod Genet 29, 617–623 (2012). https://doi.org/10.1007/s10815-012-9744-8

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  • DOI: https://doi.org/10.1007/s10815-012-9744-8

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