The varied influences of cell adhesion and spreading on gene transfection of mesenchymal stem cells on a micropatterned substrate.

Abstract Transmembrane transport of exogenous genes is broadly explored due to the flourishing 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 spreading area on transfection of exogenous genes remains unclear because it has been difficult to separate the individual influence from cell adhesion area and spreading area during normal cell culture. In this study, micropatterns were prepared to separately control adhesion area and spreading area of human bone marrow-derived mesenchymal stem cells (hMSCs). Transfection efficiency of green fluorescent protein gene to hMSCs cultured on the micropatterns was compared. The cells with larger adhesion area showed higher transfection efficiency, while cell spreading area hardly affected gene transfection efficiency. Cell adhesion area had dominant influence on gene transfection. Uptake of microparticles and BrdU staining showed that cellular uptake capacity and DNA synthesis activity increased with cell adhesion area, but were not affected by cell spreading area. The different influence of cell adhesion area and spreading area on gene transfection was correlated with their influence on cellular uptake capacity, DNA synthesis activity, focal adhesion formation, cytoskeletal mechanics and mechano-signal activation. The results suggested that cell adhesion area and spreading area had different influence on gene transfection, which should provide useful information for manipulation of cell functions in gene therapy, protein modification and cell reprogramming.