Nanotechnology is an upcoming yet promising technology with respect to the development of well-established products. Nanofibers are potentially recent additions to materials in relation to tissue engineering (TE). Nanofiber-based scaffolds are being explored as scaffolds for TE applications. Electrospinning has developed into a versatile technique to fabricate polymeric nanofiber matrices, and the ability to incorporate bioactive therapeutic molecules without adversely affecting their structural integrity and biological activity using the mild electrospinning process has generated significant interest in polymeric nanofiber-based drug release patterns by changing the mode of encapsulation as well as by varying the matrix polymer. Scaffold composition and fabrication can be controlled to confirm desired properties and biofunctionalities. Interaction between the stem cells and nanofibers are crucial in a cell-scaffold matrix while using them for different TE applications. This chapter finally talks about the stem cell-nanofiber interactions in regenerative medicine and TE. © 2013 by The Institute of Electrical and Electronics Engineers, Inc.