Monocytes are versatile cells that can fulfill proinflammatory and anti‐inflammatory functions when recruited to the liver. Recruited monocytes differentiate into tissue macrophages and dendritic cells, which sample antigens and migrate to lymph nodes to elicit T‐cell responses. The signals that determine monocyte differentiation and the role of hepatic sinusoidal endothelial cells (HSECs) in this process are poorly understood. HSECs are known to modulate T‐cell activation, which led us to investigate whether transendothelial migration of monocytes across HSECs influences their phenotype and function. Subsets of blood‐derived monocytes were allowed to transmigrate across human HSECs into a collagen matrix. Most migrated cells remained in the subendothelial matrix, but ∼10% underwent spontaneous basal to apical transendothelial migration. The maturation, cytokine secretion, and T‐cell stimulatory capacity of reverse transmigrating (RT) and subendothelial (SE) monocytes were compared. SE monocytes were mainly CD16^–, whereas 75%‐80% of RT monocytes were CD16⁺. SE monocytes derived from the CD14⁺⁺CD16⁻ subset and exhibited high phagocytic activity, whereas RT monocytes originated from CD14⁺⁺CD16⁺ and CD14⁺CD16⁺⁺ monocytes, displayed an immature dendritic cell–like phenotype (CD11c^posHLA‐DR^posCD80_loCD86_lo), and expressed higher levels of chemokine (C‐C motif) receptor 8. Consistent with a dendritic cell phenotype, RT monocytes secreted inflammatory cytokines and induced antigen‐specific CD4⁺ T‐cell activation. In contrast, SE monocytes suppressed T‐cell proliferation and activation and exhibited endotoxin tolerance. Transcriptome analysis underscored the functional differences between SE and RT monocytes. Conclusions: Migration across HSECs shapes the subsequent fate of monocytes, giving rise to anergic macrophage‐like cells in tissue and the release of immunocompetent pre–dendritic cells into the circulation. (Hepatology 2016;63:233–246)