Cell-to-cell communication it is a fundamental mechanism by which unicellular and multicellular organisms maintain relevant functions as development or homeostasis. Tunneling nanotubes (TNTs) are a type of contact-mediated cell-to-cell communication defined by being membranous structures based on actin that allow the exchange of different cellular material. TNTs have been shown to have unique structural features compared with other cellular protrusions and to contain the cell adhesion molecule N-Cadherin. Here, we investigated the possible role of N-Cadherin and of its primary linker to the actin cytoskeleton, α-Catenin in regulating the formation and transfer function of TNTs. Our data indicate that N-Cadherin through its downstream effector α-Catenin is a major regulator of TNT formation, ultrastructure, as well as of their ability to transfer material to other cells.