Raman profiling of embryo culture medium to identify aneuploid and euploid embryos

Raman-based footprint profiling of spent culture media was applied to identify aneuploid and euploid embryos and validated by preimplantation genetic testing for aneuploidy analysis.

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Volume 111, Issue 4, Pages 753–762.e1

Authors:

Bo Liang, Ph.D., Yuan Gao, Ph.D., Jiabao Xu, M.Sc., Yizhi Song, Ph.D., Liming Xuan, M.S., Ting Shi, Ph.D., Ning Wang, Ph.D., Zhaoxu Hou, Ph.D., Yi-Lei Zhao, Ph.D., Wei E. Huang, Ph.D., Zi-Jiang Chen, Ph.D., M.D.

Abstract:

Objective

To develop and validate Raman metabolic footprint analysis to determine chromosome euploidy and aneuploidy in embryos fertilized in vitro.

Design

Retrospective study.

Setting

Academic hospital.

Patient(s)

Unselected assisted reproductive technology population.

Intervention(s)

To establish the analysis protocol, spent embryo culture medium samples with known genetic outcomes from 87 human embryos were collected and measured with the use of Raman spectroscopy. Individual Raman spectra were analyzed to find biologic components contributing to either euploidy or aneuploidy. To validate the protocol via machine-learning algorithms, additional 1,107 Raman spectra from 123 embryo culture media (61 euploidy and 62 aneuploidy) were analyzed.

Main Outcome Measure(s)

Raman-based footprint profiling of spent culture media and preimplantation genetic testing for aneuploidy (PGT-A).

Result(s)

Mean-centered Raman spectra and principal component analysis showed differences in the footprints of euploid and aneuploid embryos growing in culture medium. Significant differences in Raman bands associated with small RNAs and lipids were also observed. Stacking classification based on k-nearest-neighbor, random forests, and extreme-gradient-boosting algorithms achieved an overall accuracy of 95.9% in correctly assigning either euploidy or aneuploidy based on Raman spectra, which was validated by PGT-A sequencing results.

Conclusion(s)

This study suggests that chromosomal abnormalities in embryos should lead to changes of metabolic footprints in embryo growth medium that can be detected by Raman spectroscopy. The ploidy status of embryos was analyzed by means of Raman-based footprint profiling of spent culture media and was consistent with PGT-A testing performed by next-generation sequencing.


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Editorial Office, American Society for Reproductive Medicine

Fertility and Sterility® is an international journal for obstetricians, gynecologists, reproductive endocrinologists, urologists, basic scientists and others who treat and investigate problems of infertility and human reproductive disorders. The journal publishes juried original scientific articles in clinical and laboratory research relevant to reproductive endocrinology, urology, andrology, physiology, immunology, genetics, contraception, and menopause. Fertility and Sterility® encourages and supports meaningful basic and clinical research, and facilitates and promotes excellence in professional education, in the field of reproductive medicine.

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