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

Guido Carpino; Vincenzo Cardinale; Raffaele Gentile; Paolo Onori; Rossella Semeraro; Antonio Franchitto; Yunfang Wang; Daniela Bosco; Angelo Iossa; Chiara Napoletano; Alfredo Cantafora; Giuseppe D'Argenio; Marianna Nuti; Nicola Caporaso; Pasquale Berloco; Rosanna Venere; Tsunekazu Oikawa; Lola Reid; Domenico Alvaro; Eugenio Gaudio

doi:10.1016/j.jhep.2014.01.026

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

J Hepatol. 2014 Jun;60(6):1194-202. doi: 10.1016/j.jhep.2014.01.026. Epub 2014 Feb 13.

Authors

Guido Carpino¹, Vincenzo Cardinale², Raffaele Gentile², Paolo Onori³, Rossella Semeraro², Antonio Franchitto⁴, Yunfang Wang⁵, Daniela Bosco⁶, Angelo Iossa⁷, Chiara Napoletano⁶, Alfredo Cantafora⁸, Giuseppe D'Argenio⁹, Marianna Nuti⁶, Nicola Caporaso⁹, Pasquale Berloco¹⁰, Rosanna Venere², Tsunekazu Oikawa⁵, Lola Reid⁵, Domenico Alvaro¹¹, Eugenio Gaudio¹²

Affiliations

¹ Department of Movement, Human and Health Sciences, Division of Health Sciences, University of Rome "Foro Italico", Rome, Italy.
² Department of Medico-Surgical Sciences and Biotechnologies, Polo Pontino, Sapienza University of Rome, Rome, Italy.
³ Department of Biotechnological and Applied Clinical Sciences, University of L'Aquila, L'Aquila, Italy; Department of Anatomical, Histological, Forensic Medicine and Orthopedics Sciences, Sapienza University of Rome, Rome, Italy.
⁴ Department of Anatomical, Histological, Forensic Medicine and Orthopedics Sciences, Sapienza University of Rome, Rome, Italy; Eleonora Lorillard Spencer-Cenci Foundation, Rome, Italy.
⁵ Department of Cell Biology and Physiology, Program in Molecular Biology and Biotechnology, UNC School of Medicine, Chapel Hill, NC 27599, United States.
⁶ Department of Experimental Medicine, Sapienza University of Rome, Rome, Italy.
⁷ Surgical-Medical Department for Digestive Diseases, Sapienza University of Rome, Rome, Italy.
⁸ Division of Gastroenterology, Sapienza University of Rome, Rome, Italy.
⁹ Gastroenterology Unit, Department of Clinical and Experimental Medicine, Federico II University of Naples, Italy.
¹⁰ Department of General Surgery and Organ Transplantation, Sapienza University of Rome, Rome, Italy.
¹¹ Department of Medico-Surgical Sciences and Biotechnologies, Polo Pontino, Sapienza University of Rome, Rome, Italy; Eleonora Lorillard Spencer-Cenci Foundation, Rome, Italy.
¹² Department of Anatomical, Histological, Forensic Medicine and Orthopedics Sciences, Sapienza University of Rome, Rome, Italy. Electronic address: eugenio.gaudio@uniroma1.it.

PMID: 24530598
DOI: 10.1016/j.jhep.2014.01.026

Abstract

Background & aims: 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.

Methods: 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.

Results: 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.

Conclusions: Human gallbladders contain easily isolatable cells with phenotypic and biological properties of multipotent, endodermal stem cells.

Keywords: Epithelial differentiation; Gallbladder; Liver regeneration; Pancreatic islet cells; Stem cells.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Biliary Tract / cytology
Cell Differentiation
Cholelithiasis / pathology
Cholelithiasis / surgery
Disease Models, Animal
Epithelial Cells / cytology
Gallbladder / cytology*
HT29 Cells
Hepatocytes / cytology*
Humans
Immunomagnetic Separation
Islets of Langerhans / cytology
Liver Cirrhosis, Experimental / pathology
Liver Cirrhosis, Experimental / therapy*
Liver Regeneration
Mice
Multipotent Stem Cells / cytology*
Primary Cell Culture
Stem Cell Niche*
Tissue Donors

Abstract

Publication types

MeSH terms

Grants and funding