In the fabrication of 3-D complex tissues for implantation, layer-by-layer (LBL) electrospun nanofibrous scaffolds have recently received intensive interest. However, poor cell adhesion and cell expansion between the layers in an LBL stack remain important issues. In this study, we report a mussel-inspired, biomimetic approach to functionalize the surface of PCL/gelatin nanofibrous membranes coated with poly (dopamine) (PDA). Our study demonstrates that a PDA coating on electrospun PCL/gelatin nanofibers leads to a significant change in their surface properties and a higher adhesion force. Furthermore, we found that PDA coating promotes the adhesion and growth of adipose stem cells (ADSCs). In 3-D LBL stacked scaffolds, more cells survived in a PAD-coated scaffold than in a non-coated one. The PDA coating was further demonstrated to promote the osteogenic differentiation of ADSCs in LBL paper-stacking membranes. Our study suggests that PDA-coated paper-stacking nanofiber membranes present a facile and economic method for the development of 3D tissue engineering.