Abstract
The microtubule organizing center (MTOC) frequently polarizes to a position in front of the nucleus during cell migration, but recent work has shown conflicting evidence for MTOC location in migratory polarized cells. Here, we show that subcellular localization of the MTOC is modulated by extracellular matrix stiffness. In scratch wound assays as well as single cell migration of mesenchymal stem cells (MSCs) the MTOC appears randomly positioned when cells are migrating on soft matrix, whereas on stiff matrix the MTOC is in front of the nucleus. The bulk of the microtubule density is also equally likely to be in front of or behind the nucleus on soft matrix, but it is polarized in front of the nucleus on stiff matrix. This occurred during cell migration with cells in interphase. During cytokinesis, the centrosomes polarize on either side of the chromosomes even on soft matrix, with MIIB localized strongly in the cleavage furrow which depolarizes only on soft matrix as cells exit cytokinesis. When cells are immobilized on micro-patterns printed on the top of substrates of different stiffness, MIIB polarized if the matrix was sufficiently stiff similar to results with migrating cells. However, the MTOC was randomly positioned with respect to the nucleus independent of matrix stiffness. We deduce that cell migration is necessary to orient the MTOC in front of the nucleus and that matrix stiffness helps to drive cell polarization during migration. © 2016 Wiley Periodicals, Inc.