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Stem Cells International
Volume 2017, Article ID 2679518, 15 pages
Research Article

Immunoprofiling of Adult-Derived Human Liver Stem/Progenitor Cells: Impact of Hepatogenic Differentiation and Inflammation

1Institut de Recherche Expérimentale and Clinique (IREC), Laboratory of Pediatric Hepatology and Cell Therapy, Université Catholique de Louvain, 1200 Brussels, Belgium
2Institut de Biologie et de Médecine Moléculaires, Université Libre de Bruxelles, 6041 Gosselies, Belgium
3Laboratory of Cancer Biology and Molecular Immunology, Faculty of Sciences I, Lebanese University, Beirut, Lebanon
4Laboratory of Clinical Cell Therapy, Institut Jules Bordet, Université Libre de Bruxelles (ULB), Brussels, Belgium

Correspondence should be addressed to Mustapha Najimi; eb.niavuolcu@imijan.ahpatsum

Received 28 September 2016; Revised 15 February 2017; Accepted 2 March 2017; Published 11 April 2017

Academic Editor: Eva Mezey

Copyright © 2017 Hoda El-Kehdy et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

ADHLSCs were sequentially incubated with specific growth factors/cytokines and processed for the evaluation of the hepatogenic differentiation quality. A) Differentiated ADHLSC display significant morphological changes with polygonal epithelial-like shape. Pictures were taken at magnification of 200x. Presented data are representative of at least three different experiments. B) RT-PCR analysis of hepatocyte specific gene expression profile of differentiated (Diff) compared to undifferentiated ADHLSCs (Und) confirms a positive correlation with the morphological changes. Data shown are agarose gel electrophoresis of amplification products corresponding to hepatic markers: MRP2, multidrug resisting protein-2; TDO, tryptophan 2,3-dioxygenase; CYP3A4, cytochrome P450, family 3, subfamily A, polypeptide 4; GAPDH, glyceraldehyde-3-phosphate dehydrogenase is used as house-keeping control. Presented data are representative of at least three different experiments. C) Forty μg of total protein extracted from differentiated ADHLSCs and isolated hepatocytes were analyzed using western blotting. Hepatogenic differentiation was supported, by demonstrating the expression of CYP3A4 and hepatocyte nuclear factor-4 alpha (HNF4a) proteins in differentiated ADHLSC (Diff) as compared to hepatocytes (Hep). D) After the hepatogenic differentiation process, undifferentiated (U) and differentiated (D) ADHLSCs were recovered for CYP3A4 activity analysis using P450-GloTM assay a Victor3 luminometer (PerkinElmer). Data shown are the mean ± SEM of three independent experiments (T-test ∗∗∗ p< 0.001 vs undifferentiated ADHLSC)

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