Abstract
Adhesion and spreading are essential processes of anchorage dependent cells involved in regulation of cell functions. Cells interact with their extracellular matrix (ECM) resulting in different degree of adhesion and spreading. However, it is not clear whether cell adhesion or cell spreading is more important for cell functions. In this study, 10 types of isotropical micropatterns that were composed of 2 μm microdots were prepared to precisely control the adhesion area and spreading area of human mesenchymal stem cells (MSCs). The respective influence of adhesion and spreading areas on stem cell functions was investigated. Adhesion area showed more significant influences on the focal adhesion formation, binding of myosin to actin fibers, cytoskeletal organization, cellular Young’s modulus, accumulation of YAP/TAZ in nuclei, osteogenic and adipogenic differentiation of MSCs than did the spreading area. The results indicated that adhesion area rather than spreading area played more important roles in regulating cell functions. This study should provide new insight of the influence of cell adhesion and spreading on cell functions and inspire the design of biomaterials to process in an effective manner for manipulation of cell functions.
Highlights
Influence of adhesion and spreading areas to cell functions[23,24,25,26]
The results indicated that adhesion area could regulate cytoskeletal tension and activation of nuclear Yes-associated protein (YAP)/transcriptional coactivator with PDZ-binding motif (TAZ) and had dominant effect on the differentiation of mesenchymal stem cells (MSCs)
The average size of focal adhesions (FAs) increased with increase of both adhesion area and spreading area, while the total area of FAs only increased with the increase of the adhesion area rather than spreading area (Fig. 2C and Supplementary Fig. 4)
Summary
Influence of adhesion and spreading areas to cell functions[23,24,25,26]. The controversially observed phenomena require further detailed investigation to reveal the influence of cell adhesion and spreading on cell functions. How the differentiation, the most attractive point of stem cell research, is influenced by adhesion and spreading areas remains unclear. The independent influence of adhesion and spreading area on differentiation of human mesenchymal stem cells (MSCs) was investigated by using micropatterning method to precisely control cell adhesion and spreading areas. The formation of FAs and the cytoskeletal organization in the cells cultured on the micropatterns were investigated to evaluate cell adhesion and spreading state. The osteogenic and adipogenic differentiation of MSCs were investigated to show how the adhesion and spreading areas independently influenced cell fate determination
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