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Journal of Assisted Reproduction and Genetics

, Volume 31, Issue 10, pp 1369–1376 | Cite as

Effects of group culture on the development of discarded human embryos and the construction of human embryonic stem cell lines

  • Bo Sun
  • Wenzhu Yu
  • Fang Wang
  • Wenyan Song
  • Haixia Jin
  • Yingpu Sun
Stem Cell Biology

Abstract

Purpose

To explore the effect of group culture on the developmental potential of discarded embryos in in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles and establish the human embryonic stem cell lines for future research.

Methords

Fresh discarded embryos were collected from the IVF/ICSI-ET program in the reproductive medical center of the first affiliated hospital of Zhengzhou university in this study. All zygotes were individually cultured from Day 1 to Day 3. On Day 3, discard embryos were then cultured in group of 1–4 embryos per droplet (30 μl/droplets) with a constant culture medium until Day 5 or 6. Mechanical method was used to isolate the inner cell mass (ICM) of blastocyst from the embryo. Then we inoculated the ICM on feeder layer. After identification of those cells, the human embryonic stem cell lines (hESCs) were established.

Results

In this study, we collected 1,223 fresh discarded embryos and they were sequential cultured to the blastocysts (18.07 %, 221/1,223), in which good quality blastocysts were 61(4.98 %, 61/1,223). There was no significant difference in the patients. The embryos from 1PN, 2PN, 3PN were sequential cultured to the blastocyst s(39.31 %,92/234;12.87 %,64/497;13.21 %,65/492),in which good quality blastocysts was 13.6 %(32/92),2.61 %(13/64), 3.04 %(15/65).1PN embryo’s blastulation rate and quality embryo formation rate was significantly higher than the 2PN and 3PN embryos’ (P <0.05). Three embryos group cultivation has the highest blastulation rate and quality embryo formation rate (P <0.05). In total, we successfully established 4 hESCs lines.

Conclusion

The group culture of human discard embryos can improve the blastulation rate and blastocyst quality to some extent. Three embryos group cultivate is the better culture number. Human discard embryos are good source for establishment of hESCs.

Keywords

Group culture Human discard embryo Blastocyst Human embryonic stem cell 

Notes

Acknowledgments

This study was supported by the National Natural Science Foundation of China (NO.31271605) and Graduate Programs by the fund of Zhengzhou University (NO. 12Y13402).

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Bo Sun
    • 1
  • Wenzhu Yu
    • 1
  • Fang Wang
    • 1
  • Wenyan Song
    • 1
  • Haixia Jin
    • 1
  • Yingpu Sun
    • 1
  1. 1.Reproductive Medical CenterFirst Affiliated Hospital of Zhengzhou UniversityZhengzhouChina

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