Apoptosis has been documented as a frequent hurdle phenomenon that occurs in human hepatocytes during isolation, storage, infusion and after engraftment within the recipient liver parenchyma. Apoptosis is an active form of cell death that involves programmed cellular machineries leading to a progressive self-destruction of the cell. In contrary to necrosis, it can affect individual cells within a cell population. It is characterized by chronological alteration of intracellular biochemical signaling pathways followed by cellular morphological changes, DNA fragmentation, perturbation of mitochondrial membrane function and changes in the plasma membrane. These cellular alterations can be analyzed using different methodologies on adherent, suspended and in situ engrafted hepatocytes. This chapter presents a brief overview of these techniques and provides methodology for the evaluation of hepatocyte apoptosis at the structural and biochemical levels.