Metabolomic Analysis Evidences That Uterine Epithelial Cells Enhance Blastocyst Development in a Microfluidic Device

Abstract

Here we report the use of a microfluidic system to assess the differential metabolomics of murine embryos cultured with endometrial cells-conditioned media (CM). Groups of 10, 1-cell murine B6C3F1×B6D2F1 embryos were cultured in the microfluidic device. To produce CM, mouse uterine epithelial cells were cultured in potassium simplex optimized medium (KSOM) for 24 h. Media samples were collected from devices after 5 days of culture with KSOM (control) and CM, analyzed by reverse phase liquid chromatography and untargeted positive ion mode mass spectrometry analysis. Blastocyst rates were significantly higher (p < 0.05) in CM (71.8%) compared to control media (54.6%). We observed significant upregulation of 341 compounds and downregulation of 214 compounds in spent media from CM devices when compared to control. Out of these, 353 compounds were identified showing a significant increased abundance of metabolites involved in key metabolic pathways (e.g., arginine, proline and pyrimidine metabolism) in the CM group, suggesting a beneficial effect of CM on embryo development. The metabolomic study carried out in a microfluidic environment confirms our hypothesis on the potential of uterine epithelial cells to enhance blastocyst development. Further investigations are required to highlight specific pathways involved in embryo development and implantation.

Rescue of impaired blood-brain barrier in tuberous sclerosis complex patient derived neurovascular unit

“…Neural cell and vascular chamber samples were stored at -80 °C until analyzed via Liquid Chromatography-High Resolution Mass Spectrometry (LC-HRMS)-based metabolomics in the Vanderbilt Center for Innovative Technology (CIT) using previously described methods [ 11 , 42 , 55 ] Briefly, effluent samples collected from both the vascular and brain chambers of the NVU were normalized by volume to 100 uL as previously reported [ 11 ]. Metabolites were extracted with methanol/water 80:20.…”

Rescue of Impaired Blood-Brain Barrier in Tuberous Sclerosis Complex Patient Derived Neurovascular Unit

“…Neural cell and vascular chamber samples were stored at -80°C until analyzed via Liquid Chromatography-High Resolution Mass Spectrometry (LC-HRMS)-based metabolomics in the Vanderbilt Center for Innovative Technology (CIT) using previously described methods (Brown, Codreanu et al 2016, Rogers, Sobolik et al 2018, Mancini, Schrimpe-Rutledge et al 2021) Briefly, effluent samples collected from both the vascular and brain chambers of the NVU were normalized by volume to 100 uL as previously reported (Brown, Codreanu et al 2016). Metabolites were extracted with methanol/water 80:20.…”