NeoHepatocytes generated from Patients with chronic Liver Disease can be used for autologous Cell Transplantation

Recently, we have shown that injection of hepatocyte-like cells can increase the survival in rats after extended liver resection. In order to apply this technology in humans with chronic liver diseases in an autologous setting, it is mandatory to verify whether Neohepatocyte-derived cells from these patients show similar metabolic patters as NeoHeps isolate from healthy controls.

NeoHepatocytes were generated from monocytes of alcoholic patients and healthy controls and compared to human hepatocytes. We measured glucose and urea production as well as hepatocyte markers and the TGF-ß signaling pathway using PCR, Western blot and reporter assays.

The yield of monocytes from patients (2.2±0.8*10⁷ cells/ml blood) und controls (2.5±0.9*10⁷ cells/ml blood) was comparable. There were no obvious morphological differences between the 2 sets of NeoHepatocytes after 14 days of differentiation. Although albumin expression remained very low, other hepatocyte markers, e.g. cytokeratin-18, transferrin and ADH1, increased significantly. Glucose and urea production was comparable with primary hepatocytes. Fat accumulation was induced by treatment with insulin, TGF-β and ethanol in the two sets of NeoHepatocytes and primary cells. TGF-β signaling was comparable among the investigated cell types; however, expression of Smad1 and 3 was reduced (˜30 and ˜60%). Subsequently, expression of TGF-β regulated genes, e.g. CTGF, fibronectin und collagen was low.

We could successfully generate NeoHepatocytes from patients with chronic liver disease. Cell quality and function was comparable with NeoHepatocytes from healthy controls suggesting that these cells maybe useful as a possible bridge in an autologous setting while waiting of a suitable organ.