title: Empirical chemosensitivity testing in a spheroid model of ovarian cancer using a microfluidics-based multiplex platform.
abstract: The use of biomarkers to infer drug response in patients is being actively pursued, yet significant challenges with this approach, including the complicated interconnection of pathways, have limited its application. Direct empirical testing of tumor sensitivity would arguably provide a more reliable predictive value, although it has garnered little attention largely due to the technical difficulties associated with this approach. We hypothesize that the application of recently developed microtechnologies, coupled to more complex 3-dimensional cell cultures, could provide a model to address some of these issues. As a proof of concept, we developed a microfluidic device where spheroids of the serous epithelial ovarian cancer cell line TOV112D are entrapped and assayed for their chemoresponse to carboplatin and paclitaxel, two therapeutic agents routinely used for the treatment of ovarian cancer. In order to index the chemoresponse, we analyzed the spatiotemporal evolution of the mortality fraction, as judged by vital dyes and confocal microscopy, within spheroids subjected to different drug concentrations and treatment durations inside the microfluidic device. To reflect microenvironment effects, we tested the effect of exogenous extracellular matrix and serum supplementation during spheroid formation on their chemotherapeutic response. Spheroids displayed augmented chemoresistance in comparison to monolayer culturing. This resistance was further increased by the simultaneous presence of both extracellular matrix and high serum concentration during spheroid formation. Following exposure to chemotherapeutics, cell death profiles were not uniform throughout the spheroid. The highest cell death fraction was found at the center of the spheroid and the lowest at the periphery. Collectively, the results demonstrate the validity of the approach, and provide the basis for further investigation of chemotherapeutic responses in ovarian cancer using microfluidics technology. In the future, such microdevices could provide the framework to assay drug sensitivity in a timeframe suitable for clinical decision making.
authors.affiliation: Centre de recherche du Centre hospitalier de l'Université de Montréal and Institut du cancer de Montréal, Montréal, Québec H2L 4M1, Canada.
authors.forename: Tamal
authors.initials: T
authors.lastname: Das
authors.affiliation: Centre de recherche du Centre hospitalier de l'Université de Montréal and Institut du cancer de Montréal, Montréal, Québec H2L 4M1, Canada.
authors.forename: Liliane
authors.initials: L
authors.lastname: Meunier
authors.affiliation: Centre Suisse d' Electronique et de Microtechnique, CH-7302 Landquart, Switzerland.
authors.forename: Laurent
authors.initials: L
authors.lastname: Barbe
authors.affiliation: Centre de recherche du Centre hospitalier de l'Université de Montréal and Institut du cancer de Montréal, Montréal, Québec H2L 4M1, Canada ; Division of Gynecologic Oncology, Université de Montréal, Montréal, Québec H2L 4M1, Canada.
authors.forename: Diane
authors.initials: D
authors.lastname: Provencher
authors.affiliation: ARTORG Center, University of Bern, CH-3010 Bern, Switzerland.
authors.forename: Olivier
authors.initials: O
authors.lastname: Guenat
authors.affiliation: Department of Engineering Physics, École Polytechnique de Montréal, Montréal, Québec H3T 1J4, Canada.
authors.forename: Thomas
authors.initials: T
authors.lastname: Gervais
authors.affiliation: Centre de recherche du Centre hospitalier de l'Université de Montréal and Institut du cancer de Montréal, Montréal, Québec H2L 4M1, Canada ; Département de médecine, Université de Montréal, Montréal, Québec H3T 1J4, Canada.
authors.forename: Anne-Marie
authors.initials: AM
authors.lastname: Mes-Masson