Blood-Derived Stem Cells (BDSCs) Plasticity: In Vitro Hepatic Differentiation GIORGIA ALAIMO, 1 ELIANA COZZOLI, 1 GABRIELLA MARFE, 1 LUCA ESPOSITO, 2 MARCO RANALLI, 1 DALYA HMADA, 1 ANTONIO GIORDANO, 3,4 AND ALESSANDRA GAMBACURTA 1 * 1 Department of Experimental Medicine and Surgery, University of Rome ‘‘Tor Vergata’’, Rome, Italy 2 INT-CROM, ‘Pascale Foundation’, National Cancer Institute – Cancer Research Center, Avellino, Italy 3 Sbarro Institute for Cancer Research and Molecular Medicine, Center for Biotechnology, College of Science and Technology, Temple University, Philadelphia, Pennsylvania, 4 Human Pathology and Oncology Department, University of Siena, Siena, Italy The limited availability of hepatic tissue suitable for the treatment of liver disease and drug research encourages the generation of hepatic- like cells from alternative sources as support for the regenerative medicine. Human blood derived stem cells (BDSCs) express surface markers and genes characteristic of pluripotent stem cells and have the ability to differentiate into different cell types, including tissues of endodermal origin (i.e., liver). Therefore they can represent a valuable source of hepatocytes for medicine. In this investigation, we exploited a fast hepatic differentiation protocol to generate hepatocyte-like cells from human BDSCs using only hepatocyte growth factor (HGF) and fibroblast growth factor-4 (FGF-4) as growth factors. The resulting cell population exhibited hepatic cell-like morphology and it was characterized with a variety of biological endpoint analyses. Here, we demonstrate how human BDSCs can be reprogrammed in hepatocyte-like cells by morphological, functional analysis, reverse transcriptase (RT)-PCR, and Western Blot assay. This study defines a fast and easy reprogramming strategy that facilitates the differentiation of human BDSCs along a hepatic lineage and provides a framework for a helpful source in the stem cells therapy and liver disorders. J. Cell. Physiol. 228: 1249–1254, 2013. ß 2012 Wiley Periodicals, Inc. Despite the remarkable regenerative potential and tissue turnover of liver, its diseases are becoming one of the most common causes of mortality in developing countries (Fox and Roy-Chowdhury, 2004). The World Health Organization estimates that 20 million people worldwide are affected by cirrhosis and/or liver cancer, predominantly affecting among the estimated 500 million people (nearly 10% of the world population) bearing persistent viral infections like hepatitis B (HBV) or hepatitis C (HCV) (Dietreich, 2005; Rozga, 2006). Although some people with end stage liver disease can be effectively treated with orthotopic liver transplantation (OLT), considerable morbidity and mortality are associated with this treatment. In addition, a shortage of available donors and the cost of surgery have made this treatment unavailable to many patients suffering from liver diseases in most countries. As a result, thousands of patients die each year while on a waiting list for transplantation, and many more are never put on the list. In view of these shortfalls, cell-based therapy would offer a safer and readily available alternative source of treatment for patients with chronic liver diseases (Selden and Hodgson, 2004; Nussler et al., 2006; Zhou et al., 2012). However, some major limitations of this therapy are the availability of human hepatocytes (besides the storage of donor hepatocytes), the difficult large scale hepatocytes amplification and function maintenance (Ankrum and Karp, 2010). The normal source of cells for hepatocyte transplants are livers with >50% steatosis, vascular plaques or other factors which render the tissue unsuitable for whole organ transplantation (Strom et al., 1997; Fisher et al., 2000; Muraca et al., 2002; Horslen et al., 2003; Strom and Fisher, 2003). The isolation of viable and useful cells from discarded organs has made possible the small proof of concept studies in humans (Fox et al., 1998; Fisher et al., 2000; Horslen et al., 2003; Parasassi et al., 2005). A wider use of hepatocyte transplants will require alternative and more reliable sources of cells. Xenotransplants (Nagata et al., 2003), immortalized human hepatocytes (Cai et al., 2002; Wege et al., 2003; Cai et al., 2007) and stem cells or induced pluripotent stem cells (Petersen et al., 1999; Alison et al., 2000; Theise et al., 2000; Campard et al., 2008; Basma et al., 2009; Miki et al., 2009; Iwamuro et al., 2010; Ayatollahi et al., 2011; Marongiu et al., 2011; Lee et al., 2012) have been proposed as alternative sources of cells for clinical transplants, research and toxicology studies (Parolini et al., 2010). The peripheral blood derived stem cells (BDSCs) represents a promising source of cells for regenerative medicine because their ability to differentiate into several cell types (Giovani et al., 2008). Our recent in vivo studies have shown how BDSCs are able to repopulate different organs such as muscle, bone, liver, Giorgia Alaimo, Eliana Cozzoli, Gabriella Marfe, and Alessandra Gambacurta have contributed equally to this work. Contract grant sponsor: Italian Ministry of Research’ Grant. *Correspondence to: Alessandra Gambacurta, Department of Experimental Medicine and Surgery University of Rome ‘‘Tor Vergata’’ Via Montpellier 1, 00133 Rome, Italy. E-mail: gambacur@uniroma2.it Manuscript Received: 25 October 2012 Manuscript Accepted: 29 October 2012 Accepted manuscript online in Wiley Online Library (wileyonlinelibrary.com): 8 November 2012. DOI: 10.1002/jcp.24279 ORIGINAL RESEARCH ARTICLE 1249 Journal of Journal of Cellular Physiology Cellular Physiology ß 2012 WILEY PERIODICALS, INC.