Characterization of hepatic markers in human Wharton’s Jelly-derived mesenchymal stem cells Karolien Buyl a,1,⇑ , Joery De Kock a,1 , Mehdi Najar b , Laurence Lagneaux b , Steven Branson a , Vera Rogiers a,2 , Tamara Vanhaecke a,2 a Department of Toxicology, Center for Pharmaceutical Research (CePhaR), Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090 Brussels, Belgium b Laboratory of Clinical Cell Therapy, Hôpital Erasme, Université Libre de Bruxelles, 808 Route de Lennik, B-1070 Brussels, Belgium article info Article history: Received 2 November 2012 Accepted 24 June 2013 Available online xxxx Keywords: Wharton’s Jelly Umbilical cord Adult stem cells Liver development Hepatocyte abstract Stem cell technology could offer a unique tool to develop human-based in vitro liver models that are applicable for testing of potential liver toxicity early during drug development. In this context, recent research has indicated that human Wharton’s Jelly-derived mesenchymal stem cells (hWJs) represent an interesting stem cell population to develop human hepatocyte-like cells. Here, an in-depth analysis of the expression of liver-specific transcription factors and other key hepatic markers in hWJs is evaluated at both the mRNA and protein level. Our results reveal that transcription factors that are mandatory to acquire and maintain an adult hepatic phenotype (HNF4A and HNF1A), as well as adult hepatic markers (ALB, CX32, CYP1A1, CYP1A2, CYP2B6 and CYP3A4) are not expressed in hWJs with the exception of K18. On the contrary, transcription factors involved in liver development (GATA4, GATA6, SOX9 and SOX17) and liver progenitor markers (DKK1, DPP4, DSG2, CX43 and K19) were found to be highly expressed in hWJs. These findings provide additional indication that hWJs could be a promising stem cell source to generate hepatocyte-like cells necessary for the development of a functional human-based in vitro liver model. Ó 2013 Elsevier Ltd. All rights reserved. 1. Introduction Drug-induced liver toxicity, so-called DILI is one of the major causes of post-marketing drug withdrawal (Meng, 2010). Therefore it is of utmost importance to identify the toxicological profile of drug candidates as early as possible during their development (Mandenius et al., 2011). Because of economical, but also ethical and scientific considerations, there is a growing interest from phar- maceutical companies towards the application of in vitro methods and in particular human-based in vitro models in the preclinical phase of drug development (Baranczewski et al., 2006). In this con- text, stem cell technology is an emerging science that potentially can deliver functional human hepatocyte-like cells. Human adult stem cells (hASCs) are particularly an interesting source of stem cells since they overcome the ethical debate regarding the use of human embryos for the generation of human stem cells (Leeb et al., 2011). For years, hASCs were considered to be lineage-re- stricted, but over the last decade several publications showed that postnatal stem cell populations in bone marrow, skin, umbilical cord and adipose tissue can give rise to cell types different from the tissue of origin (Aurich et al., 2009; Campard et al., 2008; Sny- kers et al., 2007; Toma et al., 2001). This multi-lineage differentia- tion capacity makes hASCs indeed an attractive source for the development of human-based alternative in vitro models that ulti- mately could replace experimental animal use. However, for hASCs we have some indication that the age of the donor is one of the parameters affecting the quality of the cells in terms of proper plu- ri-/multipotency (De Kock et al., 2009; Gago et al., 2009). Therefore it seems essential to make use of the youngest adult postnatal non- 0887-2333/$ - see front matter Ó 2013 Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.tiv.2013.06.014 Abbreviations: ACTB, beta-actin; AFP, a-foetoprotein; ALB, albumin; B2M, beta- 2-microglobulin; CX, connexin; CYP, cytochrome P450; DAPI, 4 0 ,6-diamidino-2- phenylindole; DKK, dickkopf; DMEM-LG, Dulbecco’s modified eagle medium-low glucose; DPP, dipeptidyl-peptidase; DSG, desmoglein; EGF, epidermal growth factor; FBS, fetal bovine serum; FGF, fibroblast growth factor; FOX, forkhead box; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; GJ, gap junction; hADSCs, human adipose tissue-derived stromal cells; hASCs, human adult stem cells; hBMSCs, human bone marrow-derived stromal cells; HMBS, hydroxy-methylbilane synthase; HNF, hepatocyte nuclear factor; HEPs, human hepatocytes; hSKPs, human skin-derived precursor cells; hWJs, human Wharton’s Jelly-derived mesenchymal stem cells; KRT, keratin; K, keratin; MSCs, mesenchymal stem cells; ND, not significantly detected; ONECUT, onecut homeobox; PBS, phosphate buffered saline; PFA, paraformaldehyde; qPCR, quantitative real-time reverse transcriptase poly- merase chain reaction; SOX, sex determining region Y-box; UBC, ubiquitin C. ⇑ Corresponding author. Tel.: +32 2 477 45 85; fax: +32 2 477 45 82. E-mail address: karobuyl@vub.ac.be (K. Buyl). 1 Authors contributed equally to this work. 2 Senior authors contributed equally to this work. Toxicology in Vitro xxx (2013) xxx–xxx Contents lists available at SciVerse ScienceDirect Toxicology in Vitro journal homepage: www.elsevier.com/locate/toxinvit Please cite this article in press as: Buyl, K., et al. Characterization of hepatic markers in human Wharton’s Jelly-derived mesenchymal stem cells. Toxicol. in Vitro (2013), http://dx.doi.org/10.1016/j.tiv.2013.06.014