Adult-Derived Human Liver Mesenchymal-Like Cells as a Potential Progenitor Reservoir of Hepatocytes?

Abstract

†Université catholique de Louvain, Radiodiagnostic, Cliniques universitaires St-Luc, B-1200, Brussels, Belgium ‡Université catholique de Louvain, Anatomo-pathology, Cliniques universitaires St-Luc, B-1200 Brussels, BelgiumIt is currently accepted that adult tissues may develop and maintain their own stem cell pools. Because of their higher safety profile, adult stem cells may represent an ideal candidate cell source to be used for liver cell therapies. We therefore evaluated the differentiation potential of mesenchymal-like cells isolated from adult human livers. Mesenchymal-like cells were isolated from enzymatically digested adult human liver and expanded in vitro. Cell characterization was performed using flow cytometry, RT-PCR, and immunofluorescence, whereas the differentiation potential was evaluated both in vitro after incubation with specific media and in vivo after intrasplenic transplantation of uPA +/+ -SCID and SCID mice. Adult-derived human liver mesenchymal-like cells expressed both hepatic and mesenchymal markers among which albumin, CYP3A4, vimentin, and α-smooth muscle actin. In vitro differentiation studies demonstrated that these mesenchymal-like cells are preferentially determined to differentiate into hepatocyte-like cells. Ten weeks following intrasplenic transplantation into uPA +/+ -SCID mice, recipient livers showed the presence of human hepatocytic cell nodules positive for human albumin, prealbumin, and α-fetoprotein. In SCID transplanted liver mice, human hepatocyte-like cells were mostly found near vascular structures 56 days posttransplantation. In conclusion, the ability of isolated adult-derived liver mesenchymal stem-like cells to proliferate and differentiate into hepatocyte-like cells both in vitro and in vivo leads to propose them as an attractive expandable cell source for stem cell therapy in human liver diseases.Key words: Stem cells; Transplantation; SCID mice; Differentiation; Cell therapy INTRODUCTIONattention because of their proliferative potential, their ability to (trans)differentiate into mature functional hepatocytes, and their potentially lower immunogenicity. Liver cell transplantation (LCT) has emerged as a promising alternative to whole liver transplantation (17).Beside fetal liver, evidence has been accumulated to point out the presence of stem cells into the mammalian We and others have demonstrated that LCT can be beneficial in patients with inborn errors of liver metabolism adult liver. Data obtained from injured animal models have led to identify different cell types, some of them (6,8,16,30,32,34). Recently, we demonstrated in an arginino-succinate lyase deficiency patient a correlation candidate progenitor cells for liver regeneration. Studies regarding adult liver stem cells have mostly focused on between metabolic improvement, recovery of the deficient enzyme activity, and engraftment of donor hepatooval cells as well as hematopoietic and mesenchymal cells from bone marrow origin. Oval cells are localized cytes in the recipient's liver (33). However, t...

Age‐dependent glycosylation of the sodium taurocholate cotransporter polypeptide: From fetal to adult human livers

“…Hepatocyte isolation was performed by using two‐step collagenase perfusion under good manufacturing practice‐like conditions 22, 23, 24, 25. The isolated hepatocytes were stored in liquid nitrogen until use.…”

In Vitro Differentiated Adult Human Liver Progenitor Cells Display Mature Hepatic Metabolic Functions: A Potential Tool for in Vitro Pharmacotoxicological Testing

“…It also may cells/cm 2 in six-well plates coated with rat tail collagen allow and facilitate the classification of several comtype I and in vitro differentiated into hepatocyte-like pounds according to their specific induced signaling cells after sequential incubation with growth factors and pathways and tissue activities. cytokines, as previously described (28). Medium was In this work, we evaluate the main important hepatic changed every 3 days and the cells were microscopically functions that are acquired in ADHLSCs differentiated observed every 24-36 h. At the end of the differentiain vitro.…”

“…We recently isotype II from Aspergillus (Sigma); peroxidase type II lated mesenchymal stem/progenitor cell populations from horseradish (Sigma); ethylenediaminetetraacetic from healthy adult human livers (28). The cells we isoacid disodium salt dehydrate (EDTA) (Sigma); 2-dianislated from enzymatically digested healthy liver are able idine (Ucb); sodium L-lactate (Fluka); sodium pyruvate to proliferate in vitro and show a predetermination for (Sigma); potassium dihydrogen phosphate (KH 2 PO 4 ) hepatogenic differentiation but not for osteogenic or adi-(Merck); potassium chloride (KCl) (Fluka AG); sodium pogenic differentiation.…”

“…They do not express biliary cose 6-phosphate sodium salt (Sigma); lead nitrate markers such as cytokeratin 19 (CK19). In vivo, when (Acros); ammoniac sulfide solution (Sigma); phenobarintrasplenically transplanted into uPA +/+ -SCID and SCID bital (Sigma); 4′,5,7-trihydroxyflavanone 7-rhamnoglumice, the undifferentiated cells are able to migrate, to coside (Naringin) (Sigma); rat tail collagen type I (BD engraft into the recipient liver parenchyma, and to funcBiosciences); 0.05% Trypsin-EDTA 1× (Invitrogen); Ltion up to 10 weeks posttransplantation (28).…”

Stable isotopes in the diagnosis and treatment of inherited hyperammonemia

“…Recent advances in gene therapy [40,41] hepatocyte transplantation [42,43] or adult hepatic stem cell [44] transplantation are examples. With each of these technologies, restoration of ureagenesis may not be uniform across the liver.…”