Multidimensional analysis of in vitro tridimensional (3D) cultures: Computational tools development for microscopy images

MARINA CUENCA; MELINA MUÑOZ BERNART; LUCIA CANEDO; HERNAN GRECCO; CAROLINA PEREZ CASTRO

Congreso; Reunión Conjunta de Sociedades de Biociencias.; 2017

Understanding cellular processes such as tumor progression and asymmetric division requires biological models that replicate as close as possible the conditions found in living tissue. Tridimensional (3D) cell culture technologies such as spheroids and organoids provide this model systems but create new methodological challenges to efficiently characterize their growth and internal structure. In this work, we present a modular, extendable and easy to use open source Python toolbox to obtain parameters of interest in this kind of culture. Using glioblastoma multicellular spheroids in suspension and embedded in agarose, we demonstrate a pipeline of automatic segmentation of transmission or fluorescence images, recognition of cells clusters, and morphological characterization with the possibility to apply it to tridimensional reconstructions. By making this available to the scientific community, we expect that improving the characterization of 3D cell systems provides a way to better optimize and standardize culture and assess the effect of drug in screening assays.