Isolation, Characterization, and Differentiation to Hepatocyte‐Like Cells of Nonparenchymal Epithelial Cells from Adult Human Liver

Abstract

Activation and proliferation of human liver progenitor cells has been observed during acute and chronic liver diseases. Our goal was to investigate the presence of these putative progenitors in the liver of patients who underwent lobectomy for various reasons but did not show any hepatic insufficiency. Hepatic lesions were evaluated by histological analysis. Nonparenchymal epithelial (NPE) cells were isolated from samples of human liver resections located at a distance from the lesion that motivated the operation and were cultured and characterized. These cells exhibited a marked proliferative potential. They did not express the classic set of stem cell/progenitor markers (Oct-4, Rex-1, ␣-fetoprotein, CD90, c-kit, and CD34) and were faintly positive for albumin. When cultured at confluence in the presence of hepatocyte growth factor and either epidermal growth factor or fibroblast growth factor-4, they entered a differentiation process toward hepa-

Efficient human fetal liver cell isolation protocol based on vascular perfusion for liver cell-based therapy and case report on cell transplantation

“…Little is known about whether CD90/Thy-1, which has been described for adult hepatic progenitors in rodents, [63][64][65] characterizes human hepatic progenitors (as described in 1 case 50 ), although it is often not found in the human liver. 34,48,66 Nava et al 26 could not find CD90 in human fetal hepatic progenitors from the first trimester, but they did find it in cells from the second trimester that coexpressed CD90 and hepatic markers (as we found in low percentages for the late second trimester). In contrast, Fiegel et al 64 found CD90 expression during all stages of rat liver development but at higher levels only in the early developmental stages.…”

Simultaneous characterization of progenitor cell compartments in adult human liver

“…We first cultivated epithelial progenitor cells under selective conditions (17) and obtained clusters with a typical morphology that lasted for more than two months. After evaluating a large panel of markers because no single antigen had been definitely validated (30), we observed that all of the cultured cells expressed the a6 and b1 integrins, fundamental in ensuring cell adhesion and clonality of hepatic progenitor cells (1,2).…”

“…Media were changed three times a week. LEnPCs and L-EpPCs were counted at each passage and, at the first and fifth passages, characterized for the expression of various markers (1,2,16,17,(19)(20)(21).…”

“…To this aim, intrahepatic progenitor cells committed to four different lineages were first isolated, amplified, and characterized ex vivo using previously validated culture conditions and antibody staining techniques (15)(16)(17)(18). This enabled us to select an optimal panel of monoclonal antibodies that was then used to determine the relative frequencies of the four lineages and further characterize them in clinical specimens.…”

Cell Based Therapy for Chronic Liver Disease: Role of Fetal Liver Cells in Restoration of the Liver Cell Functions

“…Isolation, expansion and differentiation of adult HPCs to functional hepatocytes has been described by several workers [30,31,32], however identification of a specific marker of HPCs still remains a challenge. Clinical studies on the repopulation of the human liver by HPCs are still awaited.…”