Mesenchymal stem cells (MSCs) are a research tool to investigate fundamental biology and are candidates for use in regenerative medicine. In this context, significant efforts have been devoted to develop technologies to control stem cell fate, including the use of soluble factors in media. However, material properties offer alternative approaches that avoid the use of soluble factors. Here, a material system capable of sustaining the growth of stem cells (maintaining stemness) and of promoting highly efficient differentiation upon external stimulation is described. Poly(ethyl acrylate) induces assembly of fibronectin (FN) into nanonetworks (FN fibrillogenesis) upon simple adsorption from solutions. It is shown that these FN nanonetworks allow growth of MSCs and maintenance of stemness for long periods of time (up to 30 d) using basal media in absence of soluble factors. Additionally, the system promotes enhanced levels of differentiation when defined supplemented media are used. The study reveals the critical role of the intermediate protein layer at the material interface to control MSC fate regardless of the properties of the underlying material and it introduces a new material system as a candidate to be used in MSC niche design.