Culturomics to investigate the Endometrial Microbiome: Proof-of-Concept: Proof-of-Concept

Abstract: The microbiome of the reproductive tract has been associated with (sub)fertility and it has been suggested that dysbiosis reduces success rates and pregnancy outcomes. The endome-trial microbiome is of particular interest given the potential impact on the embryo implanta-tion. To date, all endometrial microbiome studies have applied a metagenomics approach. A se-quencing-based technique, however, has its limitations, more specifically in adequately explor-ing low-biomass settings, such as intra-uterine/endometrial samples. In this proof-of-concept study, we demonstrate the applicability of culturomics, a high-throughput culturing approach, to investigate the endometrial microbiome. Ten subfertile women undergoing diagnostic hys-teroscopy and endometrial biopsy, as part of their routine work-up at Brussels IVF, were includ-ed after their informed consent. Biopsies were used to culture microbiota for up to 30 days in multiple aerobic and anaerobic conditions. Subsequent WASPLab®-assisted culturomics enabled a standardized methodology. Matrix-assisted laser desorption/ionization–time of flight mass spectrometry (MALDI-TOF MS) or 16S rRNA sequencing was applied to identify all of bacterial and fungal isolates. Eighty-three bacterial and two fungal species were identified. The detected species were in concordance with previously published metagenomics-based endometrial mi-crobiota analyses as 77 (91%) of them belonged to previously described genera. Nevertheless, highlighting the added value of culturomics to identify most isolates at the species level, 53 (62.4%) of the identified species were described in the endometrial microbiota for the first time. This study shows the applicability and added value of WASPLab®-assisted culturomics to inves-tigate the low biomass endometrial microbiome at a species level.