Adult Human Biliary Tree Stem Cells Differentiate to β-Pancreatic Islet Cells by Treatment with a Recombinant Human Pdx1 Peptide

PLoS One

Anastasia Renzi¹  Eugenio Gaudio¹  Pasquale Bartolomeo Berloco²  Guido Carpino³  Domenico Alvaro⁴  Gaia Scafetta⁴  Vincenzo Cardinale⁴  Rosa Puca⁴  Marella Maroder⁴  Lorenzo Nevi⁴  Lola M. Reid⁵  Giorgio Federici⁶  Michele De Canio⁶  Francesca Sicilia⁶  Domenico Casa⁷  Rocco Panetta⁷ 

[1] Department of Anatomical, Histological, Forensic Medicine and Orthopedics Sciences, Sapienza University of Rome, Rome, Italy;Department of General Surgery and Organ Transplantation, Sapienza University of Rome, Rome, Italy;Department of Health Sciences, University of Rome “Foro Italico”, Rome, Italy;Department of Medico-Surgical Sciences and Biotechnologies, Sapienza University of Rome, Rome, Italy;Departments of Cell and Molecular Physiology, Program in Molecular Biology and Biotechnology, University of North Carolina School of Medicine, Chapel Hill, North Carolina, United States of America;Departments of Science and Chemical Technologies, University of Tor Vergata, Rome, Italy;Italian Federation of Juvenile Diabetes (FDG), Rome, Italy

关键词: Cell differentiation;    Insulin;    Recombinant proteins;    Glucose;    Glucagon;    Secretion;    Transcription factors;    Stem cells;   

DOI  :  10.1371/journal.pone.0134677

学科分类：医学（综合）

来源: Public Library of Science

【 摘 要 】

Generation of β-pancreatic cells represents a major goal in research. The aim of this study was to explore a protein-based strategy to induce differentiation of human biliary tree stem cells (hBTSCs) towards β-pancreatic cells. A plasmid containing the sequence of the human pancreatic and duodenal homeobox 1 (PDX1) has been expressed in E. coli. Epithelial-Cell-Adhesion-Molecule positive hBTSCs or mature human hepatocyte cell line, HepG2, were grown in medium to which Pdx1 peptide was added. Differentiation toward pancreatic islet cells were evaluated by the expression of the β-cell transcription factors, Pdx1 and musculoapo-neurotic fibrosarcoma oncogene homolog A, and of the pancreatic hormones, insulin, glucagon, and somatostatin, investigated by real time polymerase chain reaction, western blot, light microscopy and immunofluorescence. C-peptide secretion in response to high glucose was also measured. Results indicated how purified Pdx1 protein corresponding to the primary structure of the human Pdx1 by mass spectroscopy was efficiently produced in bacteria, and transduced into hBTSCs. Pdx1 exposure triggered the expression of both intermediate and mature stage β-cell differentiation markers only in hBTSCs but not in HepG2 cell line. Furthermore, hBTSCs exposed to Pdx1 showed up-regulation of insulin, glucagon and somatostatin genes and formation of 3-dimensional islet-like structures intensely positive for insulin and glucagon. Finally, Pdx1-induced islet-like structures exhibited glucose-regulated C-peptide secretion. In conclusion, the human Pdx1 is highly effective in triggering hBTSC differentiation toward functional β-pancreatic cells.

【 授权许可】

CC BY