High-throughput 3-dimensional culture of epithelial ovarian cancer cells as preclinical model of disease

Abstract

// Victoria Heredia-Soto 1, 2 , Andrés Redondo 3, 4, 5 , Alberto Berjón 1, 6 , María Miguel-Martín 1 , Esther Díaz 4 , Roberto Crespo 4 , Alicia Hernández 7 , Laura Yébenes 1, 6 , Alejandro Gallego 3 , Jaime Feliu 2, 3, 4, 5, 8 , David Hardisson 1, 5, 6, 9 and Marta Mendiola 1, 2, 9 1 Molecular Pathology and Therapeutic Targets Research Lab, Instituto de Investigación del Hospital Universitario La Paz, IdiPAZ, Hospital Universitario La Paz, HULP, Madrid 28046, Spain 2 Centro de Investigación Biomédica en Red de Cáncer, CIBERONC. Instituto de Salud Carlos III, Madrid 28029, Spain 3 Department of Medical Oncology, Hospital Universitario La Paz, HULP, Madrid 28046, Spain 4 Translational Oncology Research Lab, IdiPAZ, Hospital Universitario La Paz, HULP, Madrid 28046, Spain 5 School of Medicine, Universidad Autónoma de Madrid, UAM, Madrid 28029, Spain 6 Department of Pathology, Hospital Universitario La Paz, HULP, Madrid 28046, Spain 7 Department of Gynecology and Obstetrics, Hospital Universitario La Paz, HULP, Madrid 28046, Spain 8 Cátedra UAM-AMGEN, Universidad Autónoma de Madrid, Campus de Cantoblanco, Madrid 28049, Spain 9 Molecular Pathology Section, Instituto de Genética Molecular y Médica, INGEMM, Hospital Universitario La Paz, HULP, Madrid 28046, Spain Correspondence to: Marta Mendiola, email: marta.mendiola@idipaz.es David Hardisson, email: david.hardisson@salud.madrid.org Keywords: ovarian cancer; epithelial mesenchymal transition; 3D cell culture model Received: August 18, 2017      Accepted: March 19, 2018      Published: April 24, 2018 ABSTRACT Background: Recent reports have identified distinct genomic patterns in ovarian carcinoma, including proliferative and mesenchymal-like groups, with worse outcome. The exact mechanisms driving the onset and progression of these tumors are still poorly understood. Additionally, researchers are concerned about the correct subtype stratification of the available cell line models, and the exploration of alternatives to monolayer culture. Identification of biomarkers to stratify cell lines, characterization of important processes as epithelial-mesenchymal transition (EMT), and the use of three-dimensional (3D) cultures as alternative models could be useful for cell line classification. Methods and Results: In this work, we present a descriptive analysis of 16 commonly used ovarian cancer cell lines. We have studied their morphology in 2- and 3D culture, and their response to cisplatin, observing in the majority of them an increased resistance in 3D. We have also performed an immunohistochemical analysis for proliferation marker Ki-67, and EMT related markers to establish phenotypes. Epithelial cells tend to show higher proliferative rates, and mesenchymal cells show an increase in EMT related markers, especially when cultured in 3D conditions. Conclusions: We have stated the complex heterogeneity of ovarian cancer models, resembling primary tumors, agreeing with the argument that the cell line model for in vitro experiments must be carefully chosen. Our results also support that tridimensional culture could be a very helpful alternative in ovarian cancer research. Regarding EMT, a very important process for the development of this disease, some related biomarkers might be further characterized for their role in this disease development.