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Increased ammonium in culture medium may promote cellular apoptosis and negatively affect pluripotency of human blastocysts

  • Gynecologic Endocrinology and Reproductive Medicine
  • Published:
Archives of Gynecology and Obstetrics Aims and scope Submit manuscript

Abstract

Purpose

To determine the association between ammonium concentration in culture medium and blastocyst development and to assess the influence of increased ammonium concentration on the expression of Bax, Bcl-2 and Oct4.

Methods

A total of 254 cleavage-stage embryos were individually cultured in 30μL G2-plus medium on Day 3, and then culture media samples were collected on Day 5 for ammonium concentration determination immediately after evaluating the embryos morphology. Poor-quality blastocysts (combined score of CC) were used for gene expression analysis. The blastocyst formation rate, good-quality blastocyst rate and relative expression levels of Bax, Bcl-2 and Oct4 were analyzed.

Results

Based on receiver operating characteristic curve, the cutoff value of ammonium concentration produced by embryos was 16.07 μmol/L (AUC = 0.722, 95% CI 0.637–0.807; P = 0.000), so all embryos were assigned to two groups according to the cutoff value: normal group (< 16.07 μmol/L) and increased group (≥ 16.07 μmol/L). There was a significant difference in blastocyst formation rate (80.5% vs 59.0%, P < 0.01) between normal group and increased group, as well as for good-quality blastocyst rate (21.0% vs 3.4%, P < 0.01). A significantly higher expression level of Bax (P < 0.05) and considerably lower expression level of Oct4 (P < 0.01) were observed in increased group compared to normal group.

Conclusion

Our data demonstrated for the first time that increased ammonium concentration in culture medium may promote cellular apoptosis and negatively affect pluripotency of human blastocyst.

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Acknowledgements

The authors thank all the patients who enrolled in the study, the clinical staff, and embryologist.

Funding

The author declare that Qingxue Zhang: MerckSerono China Research Fund for Fertility Experts, Sun Yat-sen University Clinical Research 5010 Program.

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Author notes

  1. Haijing Zhao, Qi Qiu and Songbang Ou these authors made the same contribution and were co-first author.

Authors and Affiliations

  1. Reproductive Medicine Center, Department of Gynecology and Obstetrics, Sun Yat-Sen Memorial Hospital of Sun Yat-Sen University, Guangzhou, 510120, China

    Haijing Zhao, Qi Qiu, Songbang Ou, Haiyan Lin, Wenjun Wang & Qingxue Zhang

Authors
  1. Haijing Zhao
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  2. Qi Qiu
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  3. Songbang Ou
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  4. Haiyan Lin
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  5. Wenjun Wang
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  6. Qingxue Zhang
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Contributions

HZ, QQ, and SO were responsible for the study design, ammonium concentration detection, molecular experimental procedures, acquisition of data, and the original version of the article. HL and WW recruited participants. QZ supervised the study, evaluated the clinical application, and edited the final manuscript. All co-authors read and approved the final version of the article.

Corresponding author

Correspondence to Qingxue Zhang.

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The authors declare no conflict of interest.

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Zhao, H., Qiu, Q., Ou, S. et al. Increased ammonium in culture medium may promote cellular apoptosis and negatively affect pluripotency of human blastocysts. Arch Gynecol Obstet 307, 619–624 (2023). https://doi.org/10.1007/s00404-022-06844-2

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  • DOI: https://doi.org/10.1007/s00404-022-06844-2

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