Modeling Ovarian Cancer Metastasis in Peritoneal Cavity Lining: A Method To Establish an In Vitro 3D Model of the Peritoneal Lining

The peritoneum, or the membrane that lines the abdominal cavity, constitutes peritoneal mesothelial cells supported over a layer of fibrovascular connective tissue. Metastasizing ovarian cancer cells adhere to this peritoneal lining and proliferate into cancerous growths.

To model ovarian cancer metastasis, begin by taking human fibroblasts in a conical tube and mix them with collagen - an extracellular matrix protein. Plate the suspension onto a culture dish. Incubate to facilitate collagen matrix solidification to embed the fibroblasts and mimic the connective tissue of the peritoneum.

Next, add a suspension of mesothelial cells over the fibroblast-collagen layer. Incubate the plate to allow the mesothelial cells to adhere and form a monolayer on the surface of the fibroblast-collagen layer. This 3D organotypic culture now mimics the composition and architecture of an in vivo peritoneal lining.

Finally, seed this organotypic culture with ovarian cancer cells and incubate for the desired duration. The ovarian cancer cells release certain signaling molecules that alter the mesothelial layer, enhancing cancer cell adhesion and invasion, mimicking in vivo metastatic conditions.