Evidence for multipotent endodermal stem/progenitor cell populations in human gallbladder

• Published in: Journal of hepatology Vol. 60; no. 6; pp. 1194 - 1202
• Main Authors: Carpino, Guido, Cardinale, Vincenzo, Gentile, Raffaele, Onori, Paolo, Semeraro, Rossella, Franchitto, Antonio, Wang, Yunfang, Bosco, Daniela, Iossa, Angelo, Napoletano, Chiara, Cantafora, Alfredo, D’Argenio, Giuseppe, Nuti, Marianna, Caporaso, Nicola, Berloco, Pasquale, Venere, Rosanna, Oikawa, Tsunekazu, Reid, Lola, Alvaro, Domenico, Gaudio, Eugenio
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.06.2014
• Subjects: Animals; Biliary Tract - cytology; Cell Differentiation; Cholelithiasis - pathology; Cholelithiasis - surgery; Disease Models, Animal; Epithelial Cells - cytology; Epithelial differentiation; Gallbladder; Gallbladder - cytology; Gastroenterology and Hepatology; Hepatocytes - cytology; HT29 Cells; Humans; Immunomagnetic Separation; Islets of Langerhans - cytology; Liver Cirrhosis, Experimental - pathology; Liver Cirrhosis, Experimental - therapy; Liver Regeneration; Mice; Multipotent Stem Cells - cytology; Pancreatic islet cells; Primary Cell Culture; Stem Cell Niche; Stem cells; Tissue Donors; HDM-P; PAS; PCNA; EpCAM; NCAM; TAT; R-A; cGMP; Pancreatic islet cells; K; CYP3A4; SCID; HepPar-1; HDM; HDM-C; Mdr1; PBG; RT-PCR; Gallbladder; Epithelial differentiation; HDM-H; IF; hGSC; SR; CD; KM; Mrp2; hBTSCs; PDX1; ICG; OCT4A; Stem cells; FISH; hHpSC; LGR5; SOX; FC; Liver regeneration; peribiliary gland; neural cell adhesion molecule; HDM for Pancreatic islet cell differentiation; Pancreatic and duodenal homeobox 1; Fluorescence In Situ Hybridization; human biliary tree stem cells; human gallbladder stem/progenitor cells; HDM for Cholangiocyte differentiation; Multidrug resistance-associated protein 2; tyrosine aminotransferase; octamer-binding transcription factor 4 A; Severely Combined Immunodeficient; flow cytometry; Kubota’s Medium; Rokitansky-Aschoff; indocyanine green; Proliferating Cell Nuclear Antigen; Sry-related HMG box; epithelial cell adhesion molecule; cluster differentiation; Hepatocyte Paraffin 1; secretin receptor; multidrug resistance protein 1; hormonally defined medium; current good manufacturing practice; immunofluorescence; Cytochrome P450 3A4; Periodic Acid Schiff; HDM for Hepatocyte differentiation; leucine-rich repeat containing G protein-coupled receptor 5; human hepatic stem cells; real time polymerase chain reaction; keratin
• ISSN: 0168-8278; 1600-0641; 1600-0641
• DOI: 10.1016/j.jhep.2014.01.026

Multipotent stem/progenitor cells are found in peribiliary glands throughout human biliary trees and are able to generate mature cells of hepato-biliary and pancreatic endocrine lineages. The presence of endodermal stem/progenitors in human gallbladder was explored. Gallbladders were obtained from organ donors and laparoscopic surgery for symptomatic cholelithiasis. Tissues or isolated cells were characterized by immunohistochemistry and flow cytometry. EpCAM+ (Epithelial Cell Adhesion Molecule) cells were immunoselected by magnetic microbeads, plated onto plastic in self-replication conditions and subsequently transferred to distinct serum-free, hormonally defined media tailored for differentiation to specific adult fates. In vivo studies were conducted in an experimental model of liver cirrhosis. The gallbladder does not have peribiliary glands, but it has stem/progenitors organized instead in mucosal crypts. Most of these can be isolated by immune-selection for EpCAM. Approximately 10% of EpCAM+ cells in situ and of immunoselected EpCAM+ cells co-expressed multiple pluripotency genes and various stem cell markers; other EpCAM+ cells qualified as progenitors. Single EpCAM+ cells demonstrated clonogenic expansion ex vivo with maintenance of stemness in self-replication conditions. Freshly isolated or cultured EpCAM+ cells could be differentiated to multiple, distinct adult fates: cords of albumin-secreting hepatocytes, branching ducts of secretin receptor+ cholangiocytes, or glucose-responsive, insulin/glucagon-secreting neoislets. EpCAM+ cells transplanted in vivo in immune-compromised hosts gave rise to human albumin-producing hepatocytes and to human Cytokeratin7+ cholangiocytes occurring in higher numbers when transplanted in cirrhotic mice. Human gallbladders contain easily isolatable cells with phenotypic and biological properties of multipotent, endodermal stem cells.