Stem cell therapy offers promising solutions to diseases and injuries that traditional medicines and therapies can't effectively cure. To get and explain their full therapeutic potentials, the survival, viability, integration, homing, and differentiation of stem cells after transplant must be clearly understood. To meet these urgent needs, noninvasive stem cell imaging and tracking technologies have been developed. Metabolic labeling technique is one of the most powerful tools for live cell imaging and tracking. In addition, it has many advantages for in vivo live cell imaging and tracking such as low background, correlation of survival, and very toxic and nontoxic by-products. Herein, we described the fundamental information and process of metabolic labeling techniques and suggested optimal condition for in vitro and in vivo imaging and tracking of human umbilical cord blood-derived endothelial progenitor cells (hUCB-EPCs). Based on this study, metabolic labeling techniques can be helpful for understanding the safety and effectiveness of stem cell-based therapy and determining the utility of stem cells in downstream experiments.