Over the past two decades, poly(N-isopropyl acrylamide) (pNIPAM) has become widely used for bioengineering applications. In particular, pNIPAM substrates have been used for the non-destructive release of biological cells and proteins. In this work, we review the applications for which pNIPAM substrates have been used to release biological cells, including for the study of the extracellular matrix (ECM), for cell sheet engineering and tissue transplantation, the formation of tumor-like spheroids, the study of bioadhesion and bioadsorption, and the manipulation or deformation of individual cells. The work reviewed includes that of our own group, as well as from those performing research in the field world-wide.