Mesenchymal stem cell-derived extracellular vesicles may be leveraged to treat preeclampsia — ASN Events
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Mesenchymal stem cell-derived extracellular vesicles may be leveraged to treat preeclampsia (#504)

Ashley Bannister 1 , Claire Richards 1 2 , Lin Ding 3 , Payar Radfar 3 , Sahar Ghorbanpour 4 , Louise Cole 5 , 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. Smart MCs, Sydney, NSW, Australia
  4. Institute of Health Care Engineering, School of Biomedical Engineering, Graz University of Technology, Graz, Austria
  5. Microbial Imaging Facility, Faculty of Science, University of Technology Sydney, Sydney, NSW, Australia

Background: Preeclampsia is a cardiovascular disorder associated with placental dysfunction, without a cure. Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) have anti-inflammatory, pro-angiogenic, and antioxidant effects. This study aimed to evaluate MSC-EVs cultivated on dissolvable microcarriers with various stiffnesses as a treatment for preeclampsia in a 3D bioprinted model of placental dysfunction.

 

Methods: MSCs were cultured on X-series (SmartMCs, Australia) or Cytodex3 (Cy3; Cytivia, USA) microcarriers for 3 days. EVs were isolated from 2mL conditioned media by serial ultracentrifugation. Bioprinted ACH-3P organoids were treated with hypoxic and inflammatory stimuli ± 15μg of MSC-EVs X1Medium and Cy3 from day 7. After 12 days in culture, organoids were retrieved from their matrix and immunolabelled for β-hCG and imaged using confocal microscopy to assess differentiation. Additionally, organoids were stained in situ and imaged using a widefield fluorescence microscope to assess viability. Finally, EVs were assessed for their size, concentration, and protein content using NTA, BCA assay, and LC-MS/MS proteomics.

 

Results: NTA showed X1Hard MSC-EVs produced the most EVs at 2.13 x 109 ± 1.36 x 108 particles/mL. Whereas, BCA assay revealed X1Medium MSC-EVs contained the most protein at 94 ± 4.49 µg/mL. MSC-EVs had no effect on organoid number or differentiation. However, viability staining revealed that organoids had reduced viability when exposed to inflammatory stimuli, which was not rescued by X1Medium or Cy3 EVs. Interestingly, LC-MS/MS proteomics revealed that MSC-EVs produced with X1Soft microcarriers had the most diverse proteome, containing 65 unique proteins, involving pathways related to immunomodulation and extracellular matrix organization.

 

Conclusion: This study suggests that MSC-EV production and cargo can be improved with the use of tunable microcarriers to better target preeclampsia. Future work will employ advanced imaging techniques such as fluorescence in situ hybridization and forster resonance energy transfer to assess the impacts and uptake mechanisms of MSC-EVs on placental organoids.