The inhibition of integrin internalization blocked the neural lineage specification of BMMSCs on soft substrate. Soft substrate markedly enhanced the internalization of integrin, and this internalization was mediated mainly through caveolae/raft-dependent endocytosis. In contrast, however, the level of cell surface integrin on soft substrate was significantly lower than that on stiff substrate. By using an antibody specifically recognizing the active conformation of β1 integrin, we observed that β1 integrin activation in bone marrow mesenchymal stem cells (BMMSCs) was induced by soft substrate to a significantly greater degree than by stiff substrate. Integrins are well-documented mechanosensors that are positioned at the beginning of the sensing pathway. The mechanism by which ECM elasticity induces lineage specification of stem cells has not been clearly understood.
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