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2022-RA-717-ESGO Establishment of peritoneal dECM scaffolds for 3D culture of ovarian cancer organoids
  1. Christiane Helgestad Gjerde1,
  2. Anika Langer2,
  3. Gorka Ruiz de Garibay Ponce3,
  4. Katrin Kleinmanns1,
  5. Harsh Nitin Dongre4,
  6. Rammah Elnour Mustafa4,
  7. Constantin Berger5,
  8. Robert Willoughby1,
  9. Elvira García de Jalón1,
  10. Okan Gultekin6,
  11. Erdem Bozluolcay6,
  12. Ben Davidson7,8,
  13. Pascal Gelebart1,
  14. Daniela Elena Costea4,
  15. Kaisa Lehti6,9,
  16. Emmet McCormack1,10,11 and
  17. Line Bjørge1,12
  1. 1Centre for Cancer Biomarkers CCBIO, Department of Clinical Science, University of Bergen, Bergen, Norway
  2. 2AvenCell, Dresden, Germany
  3. 3Thoracic and Clinical Translational Oncology, Sanitaria Hospital 12 de Octubre, Madrid, Spain
  4. 4Centre for Cancer Biomarkers CCBIO, Department of Clinical Medicine, University of Bergen, Bergen, Norway
  5. 5Chair Tissue Engineering and Regenerative Medicine, University Hospital Würzburg, Würzburg, Germany
  6. 6Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden
  7. 7Department of Pathology, Oslo University Hospital, Norwegian Radium Hospital, Oslo, Norway
  8. 8Factulty of Medicine, Institute of Clinical Medicine, University of Oslo, Oslo, Norway
  9. 9Department of Biomedical Laboratory Science, Norwegian University of Science and Technology, Trondheim, Norway
  10. 10Centre for Pharmacy, Department of Clinical Science, University of Bergen, Bergen, Norway
  11. 11Vivarium, Department of Clinical Science, University of Bergen, Bergen, Norway
  12. 12Department of Obstetrics and Gynecology, Haukeland University Hospital, Bergen, Norway


Introduction/Background Recently, ovarian cancer organoids have been developed, showing promising advantages compared to traditional 2D cell culture and mouse models. Organoids are 3D cell cultures and conventionally cancer cells are embedded in a gel composed of extracellular matrix (ECM) proteins. These gels do not fully mimic the native ECM of a human tumour. Natural ECM scaffolds can be generated by decellularization of different tissues (dECM). The aim of this study is to generate and characterize peritoneal extracellular matrix (PerMa) scaffold and compare to already established small intestinal submucosal scaffold (SIS). The PerMa scaffold will be used in the establishment of an ovarian cancer organoid platform.

Methodology A protocol for decellularization of porcine and human peritoneum was developed. The permeability of the scaffolds was assessed with diffusion assay. Multiphoton microscopy and rheological analyses were done to assess the collagen structure and biophysical properties of the scaffolds. Cell cultures of ovarian cancer cell lines and primary patient cells were set up.

Abstract 2022-RA-717-ESGO Figure 1

Results The decellularization was validated with histology and DNA quantification. Cell cultures were successfully established with ovarian cancer and fibroblast cell lines and primary patient cells. Growth characteristics differed significantly on PerMa and SIS. We went on to investigate whether there are structural or biophysical differences that might explain this. We found no differences in the permeability to low (4 kDa) or high (40 kDa) molecular weight molecules between SIS and PerMa, but multiphoton microscopy revealed different organization of collagen fibres. Further, rheological analyses showed differences in elasticity (storage modulus, G´) and viscosity (loss modulus, G´´).

Conclusion We have established and characterized a 3D model of ovarian cancer that better represents the tumour microenvironment. In the future we will use this model system to establish patient-derived ovarian cancer organoids with potential application for tumour biology research and personalized medicine.

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