Autologous NeoHep Derived From Chronic Hepatitis B Virus Patients' Blood Monocytes by Upregulation of cMET Signaling.

: In view of the escalating need for autologous cell-based therapy for treatment of liver diseases, a novel candidate has been explored in the present study. The monocytes isolated from hepatitis B surface antigen (HBsAg) nucleic acid test (NAT)-positive (HNP) blood were differentiated to hepatocyte-like cells (NeoHep) in vitro by a two-step culture procedure. The excess neutrophils present in HNP blood were removed before setting up the culture. In the first step of culture, apoptotic cells were depleted and genes involved in hypoxia were induced, which was followed by the upregulation of genes involved in the c-MET signaling pathway in the second step. The NeoHep were void of hepatitis B virus and showed expression of albumin, connexin 32, hepatocyte nuclear factor 4-α, and functions such as albumin secretion and cytochrome P450 enzyme-mediated detoxification of xenobiotics. The engraftment of NeoHep derived from HBsAg-NAT-positive blood monocytes in partially hepatectomized NOD.CB17-Prkdc(scid)/J mice liver and the subsequent secretion of human albumin and clotting factor VII activity in serum make NeoHep a promising candidate for cell-based therapy.

SIGNIFICANCE: This is the first report in which normal hepatocyte-like cells have been generated from blood monocytes of hepatitis B virus-infected patients without the introduction of any exogenous genetic material. These monocyte-derived hepatocyte-like cells possess cytochrome P450 enzyme cascade, suggesting their potential application in a drug-screening system. Most important, it opens up possibilities for the autologous hepatocyte-like cell transplantation to support liver functions during the acute conditions of viral hepatitis.