Dynamic three dimensional in vitro model for investigation of ovarian carcinoma progression

Ovarian carcinoma (OC) presents a significant challenge in designing effective drug treatment and in understanding cellular processes during disease progression, especially during the transition from a solid form of the tumor to a detached spheroid form, in effusion. Emerging evidence shows the various functions of sphingolipids, and specifically, Sphingolipid-1-phosphate (S1P) as regulators of cancer progression. Herein, we ventured to construct a 3D OC in vitro culture system using alginate macroporous-based scaffolds and dynamic perfusion bioreactor 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.

433 OC cells were cultured in 4 different culture forms for 3 days: (a) monolayer (b) seeded into alginate porous scaffolds, under static conditions, (c) alginate scaffolds in perfusion bioreactor, (D) cell spheroids. mRNA expression levels of S1P receptors were analyzed by RT-PCR analysis after 3 days’ culture and compared to the basal expression levels of over 250 samples from OC patients.

Cultivation of 433 cells within alginate scaffolds, cultured under flow velocity of 50 mL/h resulted in S1P receptor mRNA expression levels similar to those of primary samples from OC patients. Moreover, the relative proportions of each receptor were also similar to those of the primary tumor samples. By contrast, no such similarity was detected for monolayer and spheroid cultures.

To conclude, a novel in vitro model was designed and established for Primary OC, appropriated for experiments with a relatively large number of the samples.