Calcification, defined as mineral formation in tissue matrix, is one of the leading mechanisms in the homeostasis of bone and de novo bone formation during bone regeneration. Diminished calcification capability in bone tissue can increase the risk of bone fractures or lead to musculoskeletal disorders such as osteoporosis. A challenge in studying calcification processes is recapitulating the biological and structural aspects of tissue calcification in the research lab. In traditional culture, cells are influenced by a culture dish’s stiff 2D polystyrene environment, causing aberrant cell-matrix and cell-cell interactions. In particular, a physiological three-dimensional (3D) model that allows studying calcification mechanisms is inadequately established. This project integrates biology, material, and computational science expertise to develop innovative physiological calcifications models, an unexplored and promising scientific area. To that end, arrayed microwells made from thermoformed (micro)films allow the generation of spheroids, 3D tissue constructs formed by cellular self-organization, for studying tissue calcification mechanisms. Calcium (Ca2+) and inorganic phosphate (Pi) ion supplementation are used to induce rapid calcification to mimic physiological bone formation. This opens up possibilities for the generation of complex bone organoid models for regenerative medicine applications.
Keywords: MSCs, iPSCs, differentiation, spheroids, bone, calcium, phosphate
Techniques: thermoforming, cell culture, SEM, confocal microscopy, qPCR