Spent embryo culture medium metabolites are related to the in vitro attachment ability of blastocysts

Fiona D’Souza,Riccardo Talevi,Shubhashree Uppangala,Hanudatta S. Atreya,Satish Kumar Adiga,Guruprasad Kalthur,Sujith Raj Salian,Gitanjali Asampille

doi:10.1038/s41598-018-35342-2

Abstract

The metabolomic profile of an embryo culture medium can aid in the advanced prediction of embryonic developmental potential and genetic integrity. But it is not known if this technology can be used to determine the in vitro potential of inner cell mass (ICM) in adherence and proliferation. Here, we investigated the developmental potential of mouse 2-cell embryos carrying cisplatin-induced DNA lesions (IDL), beyond blastocyst stage using ICM outgrowth assay. The genetic integrity of ICM cells was determined by comet assay. The metabolic signatures of spent medium were recorded 84 hours post injection of hCG (hpi-hCG), and after 96 hours of extended in vitro culture (Ex 96) by NMR spectroscopy. We observed that blastocysts that lack the ability to adhere in vitro had an increased requirement of pyruvate (p < 0.01), lactate (p < 0.01), and were accompanied by a significant reduction of pyruvate-alanine ratio in the culture medium. We propose that the aforementioned metabolites from 84 hpi-hCG spent medium be further explored using appropriate experimental models, to prove their potential as biomarkers in the prediction of implantation ability of in vitro derived human embryos in clinical settings.

Highlights

Assisted reproduction technology (ART) is the most preferred therapeutic option for infertility; the inefficacy of presently practiced morphological evaluation criteria for embryo selection[1] have been implicated as one of the major limiting factors that has contributed to poor pregnancy rates[2]
As inner cell mass (ICM) formation serves as an in vitro marker to reveal the functional ability of embryos beyond blastocyst stage[21], this study investigated the implications of metabolic profiling of 84 hours post injection of hCG (hpi-hCG) spent embryo culture medium in predicting functional competence of peri-implantation stage embryos, using in vitro adherence and ICM proliferation ability
The induction of DNA lesions at the 2-cell stage resulted in a significant reduction in blastocyst formation (Control 64.5%, N = 253/392 vs induced DNA lesions (IDL) 48%, N = 325/675, p < 0.001) and hatching rate (Control 16.58%, N = 65/392 vs IDL 8.15%, N = 55/675, p < 0.001) on 108 hpi-hCG, even though the blastocysts from IDL group did not show any signs of morphological anomalies such as fragmentation and granulation (Table S1)

Summary

Introduction

Assisted reproduction technology (ART) is the most preferred therapeutic option for infertility; the inefficacy of presently practiced morphological evaluation criteria for embryo selection[1] have been implicated as one of the major limiting factors that has contributed to poor pregnancy rates[2]. In a similar retrospective approach, we were earlier able to demonstrate that biomarkers from 84 hours post injection of hCG (hpi-hCG) spent embryo culture medium could be used to predict the genetic integrity and blastocyst development 24 h in advance, using Nuclear Magnetic Resonance (NMR) spectroscopy[8]. These observations were limited to the developmental competence and genetic integrity of embryos until the blastocyst stage. As ICM formation serves as an in vitro marker to reveal the functional ability of embryos beyond blastocyst stage[21], this study investigated the implications of metabolic profiling of 84 hpi-hCG spent embryo culture medium in predicting functional competence of peri-implantation stage embryos, using in vitro adherence and ICM proliferation ability

Methods

Results

Conclusion