title: Protective effect of mesenchymal stem cell-conditioned medium on hepatic cell apoptosis after acute liver injury.
abstract: The aim of this study was to investigate the role of Mesenchymal Stem Cell (MSC) conditioned medium (CM(MSC)) 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). CM(MSC) 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. CM(MSC) were examined by flow cytometry to detect MSC-derived annexin V- and CD54/CD44-positive microparticles (MPs). In the CCl4-CM(MSC) 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-CM(MSC) treated hepatocytes at day 3 post plating. Recombinant IL-6 was induced FGL1 expression in hepatocytes derived from CCl4-treated mice suggesting that CM(MSC), which is enriched also in microparticles, attenuates CCl4-induced early apoptosis in hepatocytes through activation of FGL1.
authors: Gene and Cell Therapy Center, Hematology-BMT Unit, G. Papanikolaou Hospital, Thessaloniki, 57001, Greece. cbb.gpapanikolaou@n3.syzefxis.gov.gr
authors: Angeliki
authors: A
authors: Xagorari
authors: Eleni
authors: E
authors: Siotou
authors: Minas
authors: M
authors: Yiangou
authors: Eleftheria
authors: E
authors: Tsolaki
authors: Dimitris
authors: D
authors: Bougiouklis
authors: Leonidas
authors: L
authors: Sakkas
authors: Athanassios
authors: A
authors: Fassas
authors: Achilles
authors: A
authors: Anagnostopoulos
mesh_terms: D000208:Acute Disease; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002251:Carbon Tetrachloride / Q000633:toxicity; D056486:Chemical and Drug Induced Liver Injury / Q000517:prevention & control*; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D005340:Fibrinogen / Q000096:biosynthesis; D005434:Flow Cytometry; D022781:Hepatocytes / Q000187:drug effects*; D015850:Interleukin-6 / Q000378:metabolism; D008297:Male; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D059467:Transcriptome
chemical_list: D017077:Culture Media, Conditioned; C476403:Fgl1 protein, mouse; D015850:Interleukin-6; D005340:Fibrinogen; D002251:Carbon Tetrachloride
keywords: FGL1; Hepatic cell apoptosis; IL-6; mesenchymal stem cell-conditioned medium; microparticles
references: PLoS One. 2007 Sep 26;2(9):e941
references: 17895982
references: N Engl J Med. 1967 Oct 5;277(14):738-46 concl
references: 5342856
references: J Cell Mol Med. 2010 Jun;14(6B):1605-18
references: 19650833
references: Am J Pathol. 1998 Aug;153(2):515-25
references: 9708811
references: Toxicol Appl Pharmacol. 2009 Aug 15;239(1):55-63
references: 19481104
references: Gut. 2010 Jun;59(6):817-26
references: 19880967
references: Nucleic Acids Res. 2010 Jan;38(1):215-24
references: 19850715
references: Exp Hematol. 2005 Jan;33(1):108-19
references: 15661404
references: J Cell Physiol. 2007 Nov;213(2):286-300
references: 17559071
references: J Exp Med. 1995 Nov 1;182(5):1545-56
references: 7595224
references: Cell Res. 2002 Dec;12(5-6):353-61
references: 12528893
references: Stem Cells. 2009 Apr;27(4):971-9
references: 19353525
references: J Clin Invest. 2003 Jan;111(1):129-39
references: 12511596
references: J Cell Biochem. 2006 Aug 1;98(5):1076-84
references: 16619257
references: Am J Physiol Lung Cell Mol Physiol. 2009 Nov;297(5):L1002-11
references: 19734317
references: In Vitro. 1981 Oct;17(10):913-25
references: 6273298
references: Hepatology. 2008 May;47(5):1634-43
references: 18395843
references: Organogenesis. 2011 Apr-Jun;7(2):105-15
references: 21572253
references: Surgery. 2009 Aug;146(2):190-7
references: 19628073
references: PLoS One. 2008 Apr 02;3(4):e1886
references: 18382669
references: J Biol Chem. 2001 Jul 13;276(28):26605-13
references: 11349125
references: J Hepatol. 2005 Apr;42(4):548-56
references: 15763341
references: Stem Cells. 2003;21(5):527-35
references: 12968107
references: Proc Natl Acad Sci U S A. 2007 Jun 26;104(26):11002-7
references: 17569781
references: J Biol Chem. 2009 May 15;284(20):13335-13347
references: 19304666
references: Stem Cells. 2006 Feb;24(2):462-71
references: 16109759
references: J Biol Chem. 2003 Mar 28;278(13):11281-8
references: 12509437
references: Nat Rev Nephrol. 2010 Mar;6(3):179-83
references: 20186233
references: Biochim Biophys Acta. 2000 Jun 21;1492(1):31-44
references: 11004478
references: Biochem Biophys Res Commun. 2008 Jan 25;365(4):729-34
references: 18039467
references: FEBS Lett. 2002 Dec 18;532(3):391-5
references: 12482598