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Isolated Hepatic perfusion

Khaled A Shibany, Sabine Tötemeyer, Stefanie L Pratt, Stuart W Paine
Despite reports of the successful isolation of primary equine hepatocytes, there are no published data regarding the successful cryopreservation of these isolated cells. In this study, a detailed description of the procedures for isolation, cryopreservation, and recovery of equine hepatocytes are presented. Furthermore, the intrinsic clearance (Clint) and production of metabolites for three drugs were compared between freshly isolated and recovered cryopreserved hepatocytes. Primary equine hepatocytes were isolated using a two-step collagenase perfusion method, with an average cell yield of 2...
October 2016: Pharmacology Research & Perspectives
A Najibi, R Heidari, J Zarifi, A Jamshidzadeh, N Firoozabadi, H Niknahad
Background: Trazodone is an antidepressant agent widely administered for the treatment of depressive disorders. On the other hand, several cases of hepatic injury have been reported after Trazodone administration. Although the precise mechanism(s) of trazodone-induced liver injury is not known, some investigations proposed the role of reactive intermediates in this complication. This study was designed to investigate the role of reactive metabolites in hepatocytes injury induced by trazodone. Methods: Isolated rat hepatocytes were prepared by the method of collagenase enzyme perfusion via the portal vein...
September 19, 2016: Drug Research
Laura Hargrove, Allyson Graf-Eaton, Lindsey Kennedy, Jennifer Demieville, Jennifer Owens, Kyle Hodges, Brittany Ladd, Heather Francis
Mast cells (MCs) are immune cells that release histamine and other mediators. MC number increases after bile duct ligation (BDL) and blocking mast cell-derived histamine decreases biliary proliferation. We aimed to isolate and characterize MCs from cholestatic livers. Rats were subjected to BDL starting at 6 h and up to 14 days. MC infiltration was evaluated by toluidine blue. BDL rats were perfused using standard collagenase perfusion. Following enzymatic digestion, tissue was passed through a fine gauge needle...
August 22, 2016: Laboratory Investigation; a Journal of Technical Methods and Pathology
Eleonora M de Leede, Mark C Burgmans, Ellen Kapiteijn, Gre P M Luyten, Martine J Jager, Fred G J Tijl, Henk H Hartgrink, Dirk J Grünhagen, Joost Rothbarth, Cornelis J H van de Velde, Cornelis Verhoef, Alexander L Vahrmeijer
Uveal melanoma patients have a poor survival after the diagnosis of metastatic disease. Isolated hepatic perfusion (IHP) was developed to treat patients with unresectable metastases confined to the liver. This retrospective analysis focuses on treatment characteristics, complications, toxicity and survival after IHP. Patients with uveal melanoma metastases confined to the liver treated with IHP in two experienced hepato-pancreatic-biliary surgery centres (Erasmus MC Cancer Institute and Leiden University Medical Center) were included...
August 10, 2016: Melanoma Research
Eleonora M de Leede, Mark C Burgmans, Christian H Martini, Fred G J Tijl, Arian R van Erkel, Jaap Vuyk, Ellen Kapiteijn, Cornelis Verhoef, Cornelis J H van de Velde, Alexander L Vahrmeijer
Unresectable liver metastases of colorectal cancer can be treated with systemic chemotherapy, aiming to limit the disease, extend survival or turn unresectable metastases into resectable ones. Some patients however, suffer from side effects or progression under systemic treatment. For patients with metastasized uveal melanoma there are no standard systemic therapy options. For patients without extrahepatic disease, isolated liver perfusion (IHP) may enable local disease control with limited systemic side effects...
2016: Journal of Visualized Experiments: JoVE
P Roman, G Budziński, A Suszka-Świtek, A Caban, G Oczkowicz, R Wiaderkiewicz, F Ryszka, Z Smorąg, L Cierpka
BACKGROUND: Transgenic animals may serve as organ donors in human organ transplantation. However, the number of the studies addressing all doubts related to this issue is currently insufficient for the clinical application of this approach. The aim of this study was to analyze the hepatic tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) synthesis during a 24-hour cold preservation of the transgenic pig liver, depending on the type of transgenesis. MATERIALS AND METHODS: The study was carried out on wild-type and transgenic pig livers with transferred human α1,2-fucosyltransferase (FUT) and/or α-galactosidase (GAL) gene (four groups; n = 6)...
June 2016: Transplantation Proceedings
Kellie A Fay, Patrick N Fitzsimmons, Alex D Hoffman, John W Nichols
Isolated hepatocytes and liver S9 fractions have been used to collect in vitro biotransformation data for fish as a means of improving modeled estimates of chemical bioaccumulation. To date, however, there have been few direct comparisons of these 2 methods. In the present study, cryopreserved trout hepatocytes were used to measure in vitro intrinsic clearance rates for 6 polycyclic aromatic hydrocarbons (PAHs). These rates were extrapolated to estimates of in vivo intrinsic clearance and used as inputs to a well stirred liver model to predict hepatic clearance...
August 2, 2016: Environmental Toxicology and Chemistry
Cathal Harmon, Mark W Robinson, Ronan Fahey, Sarah Whelan, Diarmaid D Houlihan, Justin Geoghegan, Cliona O'Farrelly
The adult human liver is enriched with natural killer (NK) cells, accounting for 30-50% of hepatic lymphocytes, which include tissue-resident hepatic NK-cell subpopulations, distinct from peripheral blood NK cells. In murine liver, a subset of liver-resident hepatic NK cells have altered expression of the two highly related T-box transcription factors, T-bet and eomesodermin (Eomes). Here, we investigate the heterogeneity of T-bet and Eomes expression in NK cells from healthy adult human liver with a view to identifying human liver-resident populations...
September 2016: European Journal of Immunology
Zhi-Yu Wei, Bai-Ri Cui, Xun Cui, Yan-Ling Wu, Yang Fu, Li-Ping Liu, Hu-Ri Piao
In the present study, four novel series of 4-phenyl-1H-1,2,4-triazol-5(4H)-one derivatives containing triazole or piperazine moieties were designed, synthesized and evaluated for negative inotropic activity by measuring the left atrium stroke volume in isolated rabbit heart preparations. Almost all of the compounds showed an ability to moderate the cardiac workload by decreasing the heart rate and contractility. Among them, 7h was found to be the most potent with a change in stroke volume of -48.22 ± 0.36% at a concentration of 3×10(-5) mol/L (metoprolol: -9...
July 28, 2016: Chemical Biology & Drug Design
Keisuke Arai, Takumi Fukumoto, Masahiro Kido, Motofumi Tanaka, Kaori Kuramitsu, Hisoka Kinoshita, Shohei Komatsu, Daisuke Tsugawa, Sachio Terai, Taku Matsumoto, Tadahiro Goto, Sadaki Asari, Hirochika Toyama, Tetsuo Ajiki, Yonson Ku
PURPOSE: We assessed the predictive value of the preoperative neutrophil-to-lymphocyte ratio (NLR) in patients who underwent a two-stage treatment combining reductive surgery and percutaneous isolated hepatic perfusion for multiple hepatocellular carcinoma (HCC) with portal vein tumor thrombus (PVTT). METHODS: Forty-two patients underwent the two-stage treatment between January 2000 and December 2014 at Kobe University Hospital (Hyogo, Japan). The NLR was calculated from lymphocyte and neutrophil counts in the preoperative routine blood test...
July 27, 2016: Surgery Today
Jing Dong, Lingling Xia, Hefang Shen, Congwen Bian, Sujin Bao, Min Zhang, Yiqi Du, Yan Dai, Lijuan Zhao, Yuanhong Xu, Qiru Xiong, Jianjian Xu, Lili Xu
UNLABELLED: Liver transplantation is an effective approach to end-stage liver disease. Shortage of donor liver and increased waiting time for liver transplantation necessitate the development of an organ culture system by which livers can be cultured and maintained ex situ for a prolonged period of time. The aim of this work is to test whether cell culture condition in vitro could be used to culture whole livers ex situ without the use of erythrocytes. Twelve castrated male land race/farm young porcine livers were exposed to 30 min warm ischemia and 30 min cold perfusion...
2016: American Journal of Translational Research
Aijie Zhang, Yongming Jia, Qinghan Xu, Changyuan Wang, Qi Liu, Qiang Meng, Jinyong Peng, Huijun Sun, Pengyuan Sun, Xiaokui Huo, Kexin Liu
Alpha-naphthylisothiocyanate (ANIT) is a toxicant that is widely used in rodents to model human intrahepatic cholestasis. The aim of the study is to investigate whether effects of dioscin on ANIT-induced cholestasis are related to changes in expression of hepatic transporters in rats. Effects of dioscin on cholestasis were examined by histology and biochemical marker levels. The functional changes of hepatic transporters were determined by in vitro, in situ and in vivo. qRT-PCR and western blot were used to assess the expression of hepatic transporters in cholestatic rats...
August 15, 2016: Toxicology and Applied Pharmacology
Keisuke Arai, Takumi Fukumoto, Motofumi Tanaka, Kaori Kuramitsu, Masahiro Kido, Hisoka Kinoshita, Taku Matsumoto, Hirochika Toyama, Sadaki Asari, Tadahiro Goto, Tetsuo Ajiki, Yonson Ku
BACKGROUND: Although the effectiveness of perioperative adjuvant therapy in the treatment of hepatocellular carcinoma (HCC) has been investigated, the efficacy of preoperative therapy is unclear. Herein, we report a case of pathological complete response after percutaneous isolated hepatic perfusion (PIHP) for HCC involving portal vein tumor thrombosis (PVTT). CASE PRESENTATION: A 77-year-old woman was referred to our institute with a liver mass detected on a routine health screening...
December 2016: Surgical Case Reports
Shingo Shimada, Kenji Wakayama, Moto Fukai, Tsuyoshi Shimamura, Takahisa Ishikawa, Daisuke Fukumori, Maki Shibata, Kenichiro Yamashita, Taichi Kimura, Satoru Todo, Ikuroh Ohsawa, Akinobu Taketomi
Hydrogen gas reduces ischemia and reperfusion injury (IRI) in the liver and other organs. However, the precise mechanism remains elusive. We investigated whether hydrogen gas ameliorated hepatic I/R injury after cold preservation. Rat liver was subjected to 48-h cold storage in University of Wisconsin solution. The graft was reperfused with oxygenated buffer with or without hydrogen at 37° for 90 min on an isolated perfusion apparatus, comprising the H2 (+) and H2 (-) groups, respectively. In the control group (CT), grafts were reperfused immediately without preservation...
May 2, 2016: Artificial Organs
Kamal Hany Hussein, Kyung-Mee Park, Kyung-Sun Kang, Heung-Myong Woo
UNLABELLED: Whole organ decellularization is a cell removal process that creates a natural extracellular matrix for use in transplantation. A lack of an intact endothelial layer in the vascular network of decellularized organs results in blood clotting even with anti-coagulation treatment. Furthermore, shear stress caused by blood flow may affect reseeded parenchymal cells. We hypothesized that a heparin-gelatin mixture (HG) can act as an antithrombotic coating reagent and induce attachment and migration of endothelial cells (ECs) on vascular wall surfaces within decellularized livers, with subsequent parenchymal cell function enhancement...
July 1, 2016: Acta Biomaterialia
Reza Heidari, Neda Esmailie, Negar Azarpira, Asma Najibi, Hossein Niknahad
Sulfasalzine is a widely administered drug against inflammatory-based disorders in human. However several cases of liver injury are associated with its administration. There is no stabilized safe protective agent against sulfasalazine-induced liver injury. Current investigation was designed to evaluate if N-acetylcysteine (NAC) and dithioteritol (DTT) as thiol reducing agents and/or vitamins C and E as antioxidants have any protective effects against sulfasalazine-induced hepatic injury in an ex vivo model of isolated rat liver...
April 2016: Toxicological Research
Victoria Kegel, Daniela Deharde, Elisa Pfeiffer, Katrin Zeilinger, Daniel Seehofer, Georg Damm
Beside parenchymal hepatocytes, the liver consists of non-parenchymal cells (NPC) namely Kupffer cells (KC), liver endothelial cells (LEC) and hepatic Stellate cells (HSC). Two-dimensional (2D) culture of primary human hepatocyte (PHH) is still considered as the "gold standard" for in vitro testing of drug metabolism and hepatotoxicity. It is well-known that the 2D monoculture of PHH suffers from dedifferentiation and loss of function. Recently it was shown that hepatic NPC play a central role in liver (patho-) physiology and the maintenance of PHH functions...
2016: Journal of Visualized Experiments: JoVE
Haquima El Mourabit, Emilien Loeuillard, Sara Lemoinne, Axelle Cadoret, Chantal Housset
Myofibroblasts are matrix-producing cells with contractile properties, usually characterized by de novo expression of alpha-smooth muscle actin, that arise in fibrotic diseases. Hepatic stellate cells (HSCs), known as perisinusoidal cells containing auto-fluorescent vitamin A, are the major although not exclusive source of myofibroblasts in the injured liver. Portal myofibroblasts (PMFs) have been defined as liver myofibroblasts derived from cells that are distinct from HSCs and located in the portal tract...
2016: Frontiers in Physiology
Peter W Fan, Jacob Z Chen, M Allan Jaochico, Hank La, Ning Liu, Teresa Mulder, Robert T Cass, Matthew Durk, Kirsten Messick, Nicole Valle, Shannon Liu, Wendy Lee, James J Crawford, Joachim Rudolf, Lesley J Murray, S Cyrus Khojasteh, Matthew Wright
BACKGROUND: Significant under-prediction of in vivo clearance in rat was observed for a potent p21-activated kinase (PAK1) inhibitor, GNE1. OBJECTIVE: Rate-determining (rapid uptake) and rate-limiting (slow excretion) steps in systemic clearance and elimination of GNE1, respectively, were evaluated to better understand the cause of the in vitro-in vivo (IVIV) disconnect. METHODS: A series of in vivo, ex vivo, and in vitro experiments were carried out: 1) the role of organic cation transporters (Oct or Slc22a) was investigated in transporter knock-out and wild-type animals with or without 1-aminobenzotriazole (ABT) pretreatment; 2) the concentration-dependent hepatic extraction ratio was determined in isolated perfused rat liver; and 3) excreta were collected from both bile duct cannulated and non-cannulated rats after intravenous injection...
2016: Drug Metabolism Letters
Qidi Zhang, Ying Qu, Zhenghong Li, Qingqing Zhang, Mingyi Xu, Xiaobo Cai, Fei Li, Lungen Lu
There have been few reports on the simultaneous isolation of multiple liver cell populations thus far. As such, this study was aimed at establishing a protocol for the simultaneous separation of hepatocytes (HCs), hepatic stellate cells (HSCs), liver sinusoidal endothelial cells (LSECs) and Kupffer cells (KCs) from the rat liver and assessing the in vitro culture of these cells. Single-cell suspensions from the liver were obtained by ethylene glycol tetraacetic acid/collagenase perfusion. After low-speed centrifugal separation of HCs, pronase was added to the nonparenchymal cell fraction to eliminate the remaining HCs...
2016: Cells, Tissues, Organs
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