Despite improved prophylaxis and treatment, GvHD remains a major limitation to optimal allogeneic stem cell transplantation. Ex vivo selective depletion (SD) is a strategy to prevent GvHD, in which host-reactive donor lymphocytes are selectively eliminated from a PBSC allograft while useful donor immune function is preserved. The elimination of alloreactive and thereby GvHD-mediating T cells has been shown to be feasible in both pre-clinical and more recently clinical studies. However, SD techniques and the translational research needed for clinical application are still under development. Here we summarize and discuss the following aspects of the SD approach: selection of an appropriate allogeneic stimulator; the responder population; the alloresponse; methods for removal of alloreacting T cells; product testing; clinical considerations. Our review highlights the diversity of possible approaches and the need to develop different techniques for specific clinical applications.