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Int J Clin Exp Pathol 2013;6(5):831-840

Original Article
Protective effect of mesenchymal stem cell-conditioned medium on hepatic cell
apoptosis after acute liver injury

Angeliki Xagorari, Eleni Siotou, Minas Yiangou, Eleftheria Tsolaki, Dimitris Bougiouklis, Leonidas Sakkas, Athanassios Fassas, Achilles

Gene and Cell Therapy Center, Hematology-BMT Unit, G. Papanikolaou Hospital, Thessaloniki, 57001, Greece; Department of Genetics,
Development & Molecular Biology, School of Biology, Aristotle University of Thessaloniki, Greece; Department of Pathology, G. Papanikolaou
Hospital, Thessaloniki, 57001, Greece

Received February 4, 2013; Accepted March 28, 2013; Epub April 15, 2013; Published May 1, 2013

Abstract: The aim of this study was to investigate the role of Mesenchymal Stem Cell (MSC) conditioned medium (CMMSC) on apoptosis of
cultured mouse primary hepatocytes after in vivo carbon tetrachloride (CCl4)-induced acute liver injury. The acute liver injury was induced by
injecting CCl4 intraperitoneally in C57/BL6 mice. Hepatocytes were isolated by liver perfusion, cultured in a defined medium to maintain their
differentiation and characterized by reverse transcriptase polymerase chain reaction (RT-PCR) using the hepatic cell specific genes albumin,
hepatocyte nuclear factor 4 (HNF4) and cytokeratin 18 (CK18). CMMSC was generated from cultured bone marrow-derived MSCs (BM-MSCs).
BM-MSCs were positive for CD73, CD90, CD44 by flow cytometry and able to differentiate into chondrocytes, adipocytes and osteocytes.
Apoptosis was evaluated by both annexin V. CMMSC were examined by flow cytometry to detect MSC-derived annexin V- and
CD54/CD44-positive microparticles (MPs). In the CCl4-CMMSC treated hepatocytes, interleukin-6 (IL-6) was increased on the first day of culture
compared to control and CCl4 and was followed by upregulation of fibroblast-like-protein (FGL1) expression after 48 hrs. This was associated
with a significant decrease of annexin V positive CCl4-CMMSC treated hepatocytes at day 3 post plating. Recombinant IL-6 was induced FGL1
expression in hepatocytes derived from CCl4-treated mice suggesting that CMMSC, which is enriched also in microparticles, attenuates
CCl4-induced early apoptosis in hepatocytes through activation of FGL1. (IJCEP1302004).

Keywords: Hepatic cell apoptosis, mesenchymal stem cell-conditioned medium, FGL1, IL-6, microparticles

Address correspondence to: Dr. Angeliki Xagorari, Hematology-BMT Unit, G. Papanicolaou Hospital, Thessaloniki, 57010, Greece. E-mail: