A Novel Computer-Assisted Approach to evaluate Multicellular Tumor Spheroid Invasion Assay

• Published in: Scientific reports 2016-10, Vol.6 (1), p.35099, Article 35099
• Main Authors: Cisneros Castillo, Liliana R., Oancea, Andrei-Dumitru, Stüllein, Christian, Régnier-Vigouroux, Anne
• Format: Article
• Language: English
• Subjects: 13; 13/106; 631/114; 631/114/1314; Algorithms; Animals; Collagen; Computer Simulation; Dacarbazine - administration & dosage; Dacarbazine - analogs & derivatives; Enzyme Inhibitors - administration & dosage; Glioblastoma - drug therapy; Glioblastoma - pathology; Glioma - immunology; Glioma - pathology; High-Throughput Screening Assays - methods; High-Throughput Screening Assays - statistics & numerical data; Humanities and Social Sciences; Humans; Image processing; Image Processing, Computer-Assisted; Imaging, Three-Dimensional; Invasiveness; Light microscopy; Mice; Microscopy; Multicellular tumor spheroids; multidisciplinary; Neoplasm Invasiveness - diagnostic imaging; Neoplasm Invasiveness - pathology; Phosphotransferases (Alcohol Group Acceptor) - antagonists & inhibitors; Rats; Science; Segmentation; Software; Spheroids; Spheroids, Cellular - pathology; Time-Lapse Imaging - methods; Time-Lapse Imaging - statistics & numerical data; Tumor Cells, Cultured - pathology
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep35099

Multicellular tumor spheroids (MCTSs) embedded in a matrix are re-emerging as a powerful alternative to monolayer-based cultures. The primary information gained from a three-dimensional model is the invasiveness of treatment-exposed MCTSs through the acquisition of light microscopy images. The amount and complexity of the acquired data and the bias arisen by their manual analysis are disadvantages calling for an automated, high-throughput analysis. We present a universal algorithm we developed with the scope of being robust enough to handle images of various qualities and various invasion profiles. The novelty and strength of our algorithm lie in: the introduction of a multi-step segmentation flow, where each step is optimized for each specific MCTS area (core, halo, and periphery); the quantification through the density of the two-dimensional representation of a three-dimensional object. This latter offers a fine-granular differentiation of invasive profiles, facilitating a quantification independent of cell lines and experimental setups. Progression of density from the core towards the edges influences the resulting density map thus providing a measure no longer dependent on the sole area size of MCTS, but also on its invasiveness. In sum, we propose a new method in which the concept of quantification of MCTS invasion is completely re-thought.