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Micropatterning technology is an effective tool to study cell behaviors especially at a single cell level. In this study, photo-reactive poly(vinyl alcohol) was synthesized and micropatterned on cell-culture polystyrene plate surfaces by photolithography. Micropatterns with different sizes of central dots and numbers of protrusion lines were prepared to control the spreading area and protrusion degree of human mesenchymal stem cells (MSCs) and to investigate their influence on adipogenic differentiation of MSCs. Single MSCs adhered on the micropatterns and showed different spreading areas and protrusion degrees. Small spreading areas promoted adipogenic differentiation, while large spreading areas slowed down adipogenic differentiation. The influence of cell protrusion was dependent on the cell spreading area. Cell protrusion showed an evident influence on adipogenic differentiation when the micropattern …