Analysis of tumoral spheres growing in a multichamber microfluidic device

DENISE BELGOROSKY; TAMARA FERNÁNDEZ-CABADA ; ANA BELÉN PEÑAHERRERA-PAZMIÑO ; YANINA LANGLE ; ROSS BOOTH ; SHEKHAR BHANSALI ; MAXIMILIANO PÉREZ ; EIJAN ANA MARIA; BETIANA LERNER

CIUDAD AUTONOMA DE BUENOS AIRES

Congreso; Reunión Conjunta de Sociedades Biomédicas; 2017

SOCIEDAD ARGENTINA DE INVESTIGACIÓN CLÍNICA

Introduction: Lab on a Chip (LOC) farming systems have emerged as a powerful tool for single cell studies combined with a non-adherent cell culture substrate and single cell capture chips for the study of single cell derived tumor spheres. Furthermore this systems have the additional advantage of using low amounts of reagents and cells. Cancer is characterized by its cellular heterogeneity where only a small population of cancer stem cells (CSCs) are responsible for tumor metastases and recurrences. Thus, the in vitro strategy to the formation of a single cell-derived sphere is an attractive alternative to identify CSCs. The objective of this study, was to test the effectiveness of microdevices in order to analysis the heterogeneity within CSC populations and its interaction with different components of the extracellular matrix, using MB49-I bladder cancer cell line. The results show the usefulness of LOC as an effective method for quantification of CSC, through the formation of spheres under low adhesion conditions, establishing 32 MB49-I/ul as the optimum number of seeding cells. The device allowed the tracking of single cells derived spheres, observing different patterns of growth, resulting in large, medium, and small spheres, and the further identification of Oct-4 and CD44, CSC markers by immunofluorescence. Furthermore, we observed that spheres were able to grow in collagen IV, but no in matrigel. In conclusion, LOC devices have not only the already known advantages, but they are also a promising tool since they use small amounts of reagents and are under specific culture parameters. LOC devices could be considered as a novel technology to be used as a complement or replacement of traditional studies on culture plates.Keywords: Lab on a Chip; Cancer Stem Cell; Tumoral sphere; Oct-4; CD44