The topography of extracellular matrix regulates the differentiation of mesenchymal stem cells (MSCs). In particular, the effect of spreading shape or area on cellular differentiation and viability of individual MSCs cultured in the confined adhesive regions is an interesting fundamental issue. In this study, the adhesive patterns with the circularity of 0.1 or 1 and the areas of 314; 628; 1,256; or 2,512 μm2 were constructed using micropatterning technology. The expression of osteogenesis marker alkaline phosphatase and the apoptosis level of individual MSCs were measured using double fluorescent staining. Results indicated that individual MSCs confined in the small area showed an apoptotic tendency, and those in the large area might enter into osteogenesis. The branched shape with small circularity increased MSC viability but reduced their pluripotency compared with the circular shape. The expression of other osteogenesis markers, such as osteocalcin and Collagen I, confirmed that large and branched pattern promoted MSC osteogenesis. In addition, the transcriptional coactivator yes-associated protein (YAP) was transferred higher in the nuclei of the large and branched cells than other micropatterned groups. This study suggested that the spreading area and shape of individual MSCs regulate their viability and osteogenesis through the YAP pathway.
Keywords: apoptosis; mesenchymal stem cells; osteocyte; osteogenesis; topography.
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