Optimizing a Dynamic Three Dimensional in Vitro Model for Investigation of Ovarian Carcinoma Progression

Ovarian carcinoma (OC) presence different sites of malignancy; primary ovarian carcinoma, solid metastasis and ascites (effusion) in the abdomen. Transition from a solid form of the tumor to a detached cellular spheroid form, in effusion presents an especial challenge both in designing effective drug treatment and in understanding cellular processes during disease progression. There is a lack of an appropriate in vitro model of the tumor microenvironment mimicking three different sites of OC. There is a strong evidence, regarding role of Sphingolipid-1-phosphate (S1P) related genes in regulation of cancer progression. Herein, our main objective was to perform the mapping of the S1P axis involved genes and with acquired data to establish a 3D in vitro culture system of OC cells, using alginate macroporous-based scaffolds under dynamic conditions to mimic the 3D tumor microenvironment.

A multi chamber perfusion system was applied using ANSYS software designed optimal flow conditions, to ensure both homogeneity of the flow and shear stress within all scaffolds and of precise control over oxygen level, and thus, better mimicking in vivo conditions. The optimal flow direction and velocity were established using viability, proliferation tests and S1P related gene expression.

Our results indicated that cultivation within alginate porous scaffolds under specific dynamic conditions resulted in S1P receptor mRNA expression levels, similar to those of the in vivo samples, compared to monolayer, static scaffold and spheroid cultures, creating the Primary Carcinoma in vitro model using 433 cell line and Effusions model on base of the ES2 cell line culture.