Liver progenitor cells expressing HLA-E and method for obtaining the latter

The current invention concerns isolated liver progenitor cells, cell lines thereof, cell populations comprising such and compositions comprising such wherein the liver progenitor cells are HLA-E positive. In addition, the invention concerns a method of preparing these liver progenitor cells, wherein the method comprises the step of adding one or more cytokines to the cell medium of a culture of liver progenitor or stem cells

Clinical-grade human liver mesenchymal stem cells for the treatment of NASH-Fibrosis through immunomodulation

Background: Nonalcoholic steatohepatitis (NASH), a severe form of nonalcoholic liver diseases (NAFLD), is one of the prominent liver diseases worldwide. There is currently no approved drug for the treatment of NASH and liver transplantation is the only therapeutic approach for advanced NASH. Mesenchymal stem cells (MSCs) are promising candidates to modulate the pro-inflammatory and pro-fibrogenic environment of chronic liver because of their immunomodulatory properties. HepaStem, Human adult liver-derived MSCs isolated from organs unsuitable for transplantation can be GMP-manufactured, cryopreserved and reconstituted at the bedside as an off-the-shelf product. Safety and tolerability have been shown in a phase I/II clinical trial in patients with metabolic disorders. The proposed mechanism of action in NASH is a systemic hit-and-run suppression of inflammation and stellate cell activation through the secretion of several cytokines in response to the liver inflammation. Method: The secretion of pro-inflammatory and anti-inflammatory cytokines including HGF, IDO, PGE2 was measured using multiplexed immunoassays in cell culture with or without inflammation cocktail. The anti-inflammatory effect of HepaStem was investigated in co-culture systems with T-lymphocytes in a mixed leukocyte reaction as well as with immature and mature dendritic cells. In a preclinical high-fat model, the potency of HepaStem was compared to a vehicle, either with or without the use of immunosuppression (cyclosporine), to factor in the use of human cells in an animal model. Results: Secretion of anti-inflammatory and anti-fibrotic cytokines was increased with the addition of an inflammatory cytokines in the culture medium HepaStem inhibited both T-lymphocyte response and the dendritic cell generation and function in co-culture experiments. In the NASH model, while the immunosuppression by itself did not affected the disease progression, cell-based treatment (3 IV injections 12.5x106 cells/kg) significantly and dose-dependently decreased collagen deposition in the pericentral region as shown by Sirius red staining. A single HepaStem injection significantly decreased the NAS score, which was mainly attributed to reduction in inflammation and thus supporting the proposed mechanism of action. Conclusion: Our results suggest that clinical grade liver progenitor cells have anti-fibrosis and anti-NASH effects, both in vitro and in a pre-clinical NASH model. This observation provides significant evidences to open new phase I/II studies in NASH patients as well as to apply MSCs for the treatment of chronic liver disorders

GMP-manufactured, heterologous human adult-derived progenitor cell therapy for NASH-fibrosis

Human Adult Liver-derived Progenitor Cells (Hepastem) were reproducibly isolated, manufactured and cryopreserved in clinical grade conditions. A bedside cell product formulation protocol was validated, to create an off-the-shelf product, easy to apply, without the need of GMP laboratories near the patient. The safety and tolerability of their intraportal injection has been shown in a phase I/II trial in young patients with metabolic disorders (UCD and Crigler-Najjar). Thanks to their mesenchymal features, the cells also display immunomodulatory properties which makes them promising cell therapy candidates for the treatment of chronic liver diseases, like fibrosis caused by steatohepatitis. The proposed mechanism of action is a systemic hit-and-run suppression of inflammation and stellate cell activation (which causes collagen deposition) through the secretion of potent cytokines. Indeed, in vitro studies have shown a significant increase of anti-inflammatory and anti-fibrotic cytokines including HGF, IDO, and PGE2, upon their incubation with an inflammatory cocktail. Co-culture studies demonstrated that the anti-inflammatory response of those liver progenitor cell is mediated via the inhibition of both T-lymphocyte response in an MLR reaction and the generation of dendritic cells when induced and matured in vitro. In a preclinical established high-fat model, the potency of Hepastem was compared to a vehicle, either with or without immunosuppression (cyclosporine). While immunosuppression itself did not affect the disease progression in this model, Hepastem treatment (1, 2 or 3 intravenous injections of 12.5x106 cells/kg) significantly and dose-dependently decreased collagen deposition in the pericentral region, as revealed by Sirius Red staining. Interestingly, a single injection significantly decreased the NAS score, which was mainly attributed to reduction in inflammation, which confirms the proposed anti-inflammatory mechanism of action. Oil red O and glycogen contents were less obvious in the treated groups, independently of the use of immunosuppression. Altogether, these data reveal the anti-fibrosis and anti-NASH potential of Hepastem under immunosuppression regimen and support its clinical development for the treatment of late phase NASH patients showing fibrosis formation