Oral Presentation ESA-SRB-ANZOS 2025 in conjunction with ENSA

Bioprinted placental organoids as a high-throughput platform for drug screening during early pregnancy (128336)

Ashley Bannister 1 , Claire Richards 1 2 , Louise Cole 3 , Lana McClements 1 2
  1. School of Life Sciences, Faculty of Science, University of Technology Sydney, Sydney, NSW, Australia
  2. Institute for Biomedical Materials and Devices, Faculty of Science, University of Technology Sydney, Sydney, NSW, Australia
  3. Microbial Imaging Facility at the Australian Institute for Microbiology and Infection, Faculty of Science, University of Technology Sydney, Sydney, NSW, Australia

Preeclampsia is a cardiovascular condition that occurs during pregnancy, associated with placental dysfunction. Without a cure, women and children affected by preeclampsia have an increased risk of chronic health conditions later in life. 3D bioprinted models of the placenta can provide insight into placental development for therapeutic testing. We aimed to assess the effectiveness of aspirin and metformin at abrogating an inflammatory response in first trimester trophoblast organoids.

ACH-3P trophoblast cells were bioprinted in a poly-ethylene glycol (PEG)-based matrix using a RASTRUM bioprinter (Inventia Life Sciences) for 12 days. Organoids were treated with tumour necrosis factor alpha (TNFα; 20ng/mL) with or without aspirin (0.5mM) or metformin (0.5mM). Organoid size, number, and metabolic activity were assessed via live cell imaging and Alamar blue assay. On day 12, culture media was collected to assess inflammation via a C-Reactive Protein (CRP) ELISA. Finally, immunofluorescence staining was performed to assess trophoblast differentiation by confocal microscopy.

There were no statistically significant differences in organoid size or number between conditions. However, from day 9 a statistically significant decrease in metabolic activity was observed in TNF-α ± aspirin/metformin groups compared to the control (p<0.01).  Aspirin (p<0.001) or metformin (p<0.01) on their own also reduced ACH-3P organoid metabolic activity.  TNFα induced a ~25-fold increase in CRP concentration within organoid media compared to all other groups. Finally, confocal imaging revealed that organoids from all conditions were able to differentiate into extravillous trophoblasts (EVTs; HLA-G) and syncytiotrophoblasts (STBs; β-hCG). Although there were no statistically significant differences in EVT differentiation, STB differentiation was reduced in all groups (p<0.05) except TNFα + metformin (p=0.07).

In this study, we demonstrate that bioprinted ACH-3P trophoblast organoids can be used as a high-throughput model for drug testing and to determine multiple end points related to placental development in both health and disease.