A Novel Nucleus-Based Classifier for Discrimination of Osteoclasts and Mesenchymal Precursor Cells in Mouse Bone Marrow Cultures

Authors: Andreas Heindl, Alexander K. Seewald, Martin Schepelmann, Radu Rogojanu, Giovanna Bises, Theresia Thalhammer, Isabella Ellinger

Abstract:

Bone remodeling occurs by the balanced action of bone resorbing osteoclasts (OC) and bone-building osteoblasts. Increased bone resorption by excessive OC activity contributes to malignant and non-malignant diseases including osteoporosis. To study OC differentiation and function, OC formed in in vitro cultures are currently counted manually, a tedious procedure which is prone to inter-observer differences. Aiming for an automated OC-quantification system, classification of OC and precursor cells was done on fluorescence microscope images based on the distinct appearance of fluorescent nuclei. Following ellipse fitting to nuclei, a combination of eight features enabled clustering of OC and precursor cell nuclei. After evaluating different machine-learning techniques, LOGREG achieved 74% correctly classified OC and precursor cell nuclei, outperforming human experts (best expert: 55%). In combination with the automated detection of total cell areas, this system allows to measure various cell parameters and most importantly to quantify proteins involved in osteoclastogenesis.

Keywords: osteoclasts, machine learning, ellipse fitting.

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1327656

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