Understanding mechanisms responsible for engraftment of hematopoietic stem cells (HSC) is important to achieve successful HSC transplantation. Homing of HSC to the bone marrow niche is believed to be a crucial step for engraftment. However, the molecular mechanisms that regulate HSC homing are not understood well. Migration of HSC in response to cytokines and chemokines is believed to be one of the critical steps of HSC homing and mobilization. Evaluating the migration of HSC will help to understand the mechanisms responsible for their homing and/or mobilization. In this chapter, we will describe the methodology utilized in our laboratory to evaluate migration of human CD34+ cells that contain HSC.