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Updated: Mar 28, 2026
Cell-Lineage Guided Mass Spectrometry Proteomics in the Developing Frog Embryo
Published on: April 21, 2022
Early protein profile of human embryonic secretome
Carlo Foresta1, Filippo Maria Ubaldi2, Laura Rienzi2
1University of Padova, Department of Medicine & Andrology and Reproductive Medicine Unit, 35128, Padova, Italy, carlo.foresta@unipd.it.
Abstract:
Embryos obtained by in vitro fertilization are currently assessed by morphology, but displays limitations with over 70% of embryos failing to implant. In this study, we performed HPLC-MS/MS analysis on the conditioned medium obtained from 50 human embryos at the 3rd day of in vitro culture. 70 proteins were identified in the medium of 48 embryos. Validation by protein array on two pools of 11 and 9 conditioned media, showed a protein pattern overlap with HPLC-MS/MS of respectively 72% and 78%. Unsupervised hierarchical cluster analysis on protein spectra, allowed to divide embryos into 3 clusters. The first cluster selectively lacked of proteins involved in programmed cell death. The third cluster was devoid of proteins involved in cell development. Embryos taking the shortest time to develop into 5 cell morulas, featuring lower implantation rate, significantly segregated in the third cluster (P=0.047). Multiple linear regression analysis, identified 12 predictive proteins for transfer success (P < 0.05). Proteomics of embryo secretome aids in understanding embryo physiology and in improving assisted reproductive technology.
Insights
Analyzing human embryo secretomes using proteomics reveals distinct protein patterns. These patterns correlate with embryo development and implantation success, offering new insights beyond traditional morphology assessment in assisted reproductive technology.
Area of Science:
- Proteomics
- Embryology
- Assisted Reproductive Technology (ART)
Background:
- Current in vitro fertilization (IVF) embryo assessment relies on morphology, which has limitations, as over 70% of embryos fail to implant.
- There is a need for improved methods to predict embryo viability and implantation potential.
Purpose of the Study:
- To investigate the potential of embryo secretome proteomics for assessing human embryo quality.
- To identify protein biomarkers in conditioned medium associated with embryo development and implantation success.
Main Methods:
- High-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was used to analyze the conditioned medium from 50 human embryos at day 3 of in vitro culture.
- Protein identification and quantification were performed, followed by validation using protein arrays.
- Unsupervised hierarchical cluster analysis and multiple linear regression were employed to analyze protein spectra and identify predictive markers.
Main Results:
- 70 proteins were identified in the conditioned medium of 48 embryos, with high overlap between HPLC-MS/MS and protein array validation (72-78%).
- Embryos were classified into 3 clusters based on protein profiles; one lacked cell death proteins, another lacked cell development proteins.
- Embryos with faster development to the 5-cell morula stage and lower implantation rates segregated into the cluster lacking cell development proteins (P=0.047).
- Twelve predictive proteins for successful embryo transfer were identified (P < 0.05).
Conclusions:
- Proteomic analysis of the human embryo secretome provides valuable insights into embryo physiology.
- Secretome proteomics can potentially improve the prediction of embryo implantation potential, enhancing assisted reproductive technology outcomes.
- This approach offers a novel method for embryo assessment beyond morphological evaluation.
