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Transmembrane transport of exogenous genes is widely investigated because of high demand for gene therapy. Both gene carriers and cellular conditions can affect gene transfection efficiency. Although cell morphology has been reported to affect cell functions, the influence of cell adhesion area and cell spreading area on the transfection of exogenous genes remains unclear because it is difficult to separate the individual influence of these areas during normal cell culture. In this study, micropatterns were prepared to separately control the adhesion and spreading areas of human bone marrow-derived mesenchymal stem cells (hMSCs). Transfection efficiency of the green fluorescent protein gene to hMSCs cultured on the micropatterns was compared. Cells with a larger adhesion area showed higher transfection efficiency, while cell spreading area hardly affected gene transfection efficiency. Cell adhesion area …