Hepatic encephalopathy (HE) is a nervous system disorder produced by liver failure. HE patients display a broad range of symptoms, involving motor, emotional, and cognitive disorders. HE severe cases end in patient death. This disease is a worldwide affection, and which number is increasing in parallel to liver pathologies associated with occidental lifestyles. The therapeutic HE approaches are limited by liver transplantation offers and by the absence of human hepatocyte research models. To address this technical limitation, we present a method to obtain human liver spheroids from unmanipulated peripheral blood. Human liver spheroids are obtained by using blood-derived pluripotent stem cells (BD-PSCs), previously generated from unmanipulated peripheral blood by activation of human membrane-bound GPI-linked protein, following differentiation into human hepatocytes. Human liver spheroids possess better physiological responses than 2D cultures, including a more realistic biologically microenvironment, and more accurate prediction of disease state and in vivo-like drug response. This chapter provides the guidelines for discovering our method, introduces a promising new source of human hepatocyte spheroids, and offers a new technical HE research tool.