Iron metabolism is tightly regulated so iron is available for biological functions while preventing its cytotoxic effects. Macrophages play a central role in establishing this balance. Kupffer Cells (KCs), liver resident macrophages, are involved in the elimination of damaged erythrocytes, ensuring systemic iron recycling to prevent damage from excess iron deposition in organs. Moreover, the hepcidin (HAMP)/ferroportin axis is critical to coordinate cellular iron export. Hepcidin induces the degradation of the iron exporter ferroportin, suppressing iron release from macrophages and parenchymal cells. The hepatocyte-derived hepcidin control on systemic iron homeostasis is well-known, but the autocrine role of hepcidin produced by the macrophage remains to be more defined. Here, we observed that iron overloading by iron dextran injection into wild-type mice resulted in a rapid and partial loss of embryonically-derived KCs (EmKCs composing the KC pool in healthy livers). Concomitantly, monocyte-derived macrophages were recruited to the liver and acquired the prototypical KC marker CLEC2. However, engraftment of these monocyte-derived KCs was only transient, and EmKCs proliferated to replenish the KC pool later. Similar studies of iron overloading were replicated in mice with hepcidin deficiency in macrophages (Hamp^MacKO; Lysm-Cre x Hamp^lox/lox), including KCs. Decreased EmKCs density was rapidly observed in both Hamp^MacKO and Hamp^lox/lox control mice following iron dextran injection. However, Hamp^MacKO mice failed to restore their EmKCs pool overtime, and associated with a reduced proliferation rate. These preliminary observations suggest that hepcidin plays a role in Kupffer Cells response to iron overloading.