title: Recellularization of rat liver scaffolds by human liver stem cells.
abstract: In the present study, rat liver acellular scaffolds were used as biological support to guide the differentiation of human liver stem-like cells (HLSC) to hepatocytes. Once recellularized, the scaffolds were maintained for 21 days in different culture conditions to evaluate hepatocyte differentiation. HLSC lost the embryonic markers (alpha-fetoprotein, nestin, nanog, sox2, Musashi1, Oct 3/4, and pax2), increased the expression of albumin, and acquired the expression of lactate dehydrogenase and three subtypes of cytochrome P450. The presence of urea nitrogen in the culture medium confirmed their metabolic activity. In addition, cells attached to tubular remnant matrix structures expressed cytokeratin 19, CD31, and vimentin. The rat extracellular matrix (ECM) provides not only a favorable environment for differentiation of HLSC in functional hepatocytes (hepatocyte like) but also promoted the generation of some epithelial-like and endothelial-like cells. When fibroblast growth factor-epidermal growth factor or HLSC-derived conditioned medium was added to the perfusate, an improvement of survival rate was observed. The conditioned medium from HLSC potentiated also the metabolic activity of hepatocyte-like cells repopulating the acellular liver. In conclusion, HLSC have the potential, in association with the natural ECM, to generate in vitro a functional "humanized liver-like tissue."
authors: 1Translational Center for Regenerative Medicine and Molecular Biotechnology Center, University of Torino, Torino, Italy.
authors: Victor
authors: V
authors: Navarro-Tableros
authors: 1Translational Center for Regenerative Medicine and Molecular Biotechnology Center, University of Torino, Torino, Italy.
authors: Maria Beatriz
authors: MB
authors: Herrera Sanchez
authors: 1Translational Center for Regenerative Medicine and Molecular Biotechnology Center, University of Torino, Torino, Italy.
authors: Federico
authors: F
authors: Figliolini
authors: 2Liver Transplantation Center, University of Torino, Torino, Italy.
authors: Renato
authors: R
authors: Romagnoli
authors: 3EMEA LA Medical Board, Fresenius Medical Care, Bad Homburg, Germany.
authors: Ciro
authors: C
authors: Tetta
authors: 4Department of Medical Sciences, University of Torino, Torino, Italy.
authors: Giovanni
authors: G
authors: Camussi
mesh_terms: D053687:Adult Stem Cells / Q000166:cytology*; D000418:Albumins / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D017209:Apoptosis; D002448:Cell Adhesion; D002454:Cell Differentiation; D002455:Cell Division; D017077:Culture Media, Conditioned / Q000737:chemistry; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000235:genetics; D005109:Extracellular Matrix; D005326:Fetal Proteins / Q000096:biosynthesis / Q000235:genetics; D020869:Gene Expression Profiling; D022781:Hepatocytes / Q000166:cytology*; D006801:Humans; D007381:Intermediate Filament Proteins / Q000096:biosynthesis / Q000235:genetics; D007770:L-Lactate Dehydrogenase / Q000096:biosynthesis / Q000235:genetics; D008099:Liver / Q000648:ultrastructure*; D008297:Male; D009584:Nitrogen / Q000032:analysis; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D054457:Tissue Scaffolds; D014508:Urea / Q000378:metabolism
chemical_list: D000418:Albumins; D017077:Culture Media, Conditioned; D005326:Fetal Proteins; D007381:Intermediate Filament Proteins; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D014508:Urea; D003577:Cytochrome P-450 Enzyme System; D007770:L-Lactate Dehydrogenase; D009584:Nitrogen