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Experimental bio-artificial liver:Importance of the architectural design on ammonia detoxification performance

World Journal of Hepatology 2018年第10期10卷 719-730页

作者： María Dolores Pizarro María Eugenia Mamprin Lucas Damián Daurelio Joaquín Valentín Rodriguez María Gabriela Mediavilla Consejo Nacional de Investigaciones Científicas y Técnicas(CONICET) Rosario S2002 LRKArgentina Laboratorio de Investigaciones en Fisiología y Biología Molecular Vegetal(LIFiBVe) Facultad de Ciencias AgrariasUniversidad Nacional del LitoralEsperanza 3080Argentina Facultad de Ciencias Bioquímicas y Farmacéuticas Universidad Nacional de RosarioRosario S2002 LRKArgentina Centro Binacional de Criobiología Clínica y Aplicada(CAIC) Universidad Nacional de RosarioRosario S2011 BXNArgentina Instituto de Biología Molecular y Celular de Rosario(IBR CONICET-UNR)Consejo Nacional de Investigaciones Científicas y Tecnológicasy Universidad Nacional de RosarioRosario S2002 LRKArgentina

AIM To determine the influence of the construction design over the biological component's performance in an experimental bio-artificial liver(BAL) *** Two BAL models for liver microorgans(LMOs) were constructed. First, we constructed a cylindrical BAL and tested it without the biological component to establish its correct functioning. Samples of blood and biological compartment(BC) fluid were taken after 0, 60, and 120 min of perfusion. Osmolality, hematocrit, ammonia and glucose concentrations, lactate dehydrogenase(LDH) release(as a LMO viability parameter), and oxygen consumption and ammonia metabolizing capacity(as LMO functionality parameters) were determined. CPSI and OTC gene expression and function were measured. The second BAL, a "flat bottom" model, was constructed using a 25 cm2 culture flask while maintaining all other components between the models. The BC of both BALs had the same capacity(approximately 50 cm3) and both were manipulated with the same perfusion system. The performances of the two BALs were compared to show the influence of *** The cylindrical BAL showed a good exchange of fluids and metabolites between blood and the BC, reflected by the matching of osmolalities, and glucose and ammonia concentration ratios after 120 min of perfusion. No hemoconcentration was detected, the hematocrit levels remained stable during the whole study, and the minimal percentage of hemolysis(0.65% ± 0.10%) observed was due to the action of the peristaltic pump. When LMOs were used as biological component of this BAL they showed similar values to the ones obtained in a Normothermic Reoxygenation System(NRS) for almost all the parameters assayed. After 120 min, the results obtained were: LDH release(%): 14.7 ± 3.1 in the BAL and 15.5 ± 3.2 in the NRS(n = 6); oxygen consumption(μmol/min?g wet tissue): 1.16 ± 0.21 in the BAL and 0.84 ± 0.15 in the NRS(n = 6); relative expression of Cps1 and Otc: 0.63 ± 0.12 and 0.67 ± 0.20, respectively,

关键词： Bio-artificial liver Ammonia detoxification Device design Ornithine transcarbamylase Rat liver microorgans Carbamyl Phosphate SynthetaseⅠ

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Hepatocyte selection medium eliminating induced pluripotent stem cells among primary human hepatocytes

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World Journal of Methodology 2015年第3期5卷 108-114页

作者： Minoru Tomizawa Fuminobu Shinozaki Yasufumi Motoyoshi Takao Sugiyama Shigenori Yamamoto Naoki Ishige Department of Gastroenterology National Hospital Organization Shimoshizu Hospital Department of Radiology National Hospital Organization Shimoshizu Hospital Department of Neurology National Hospital Organization Shimoshizu Hospital Department of Rheumatology National Hospital Organization Shimoshizu Hospital Department of Pediatrics National Hospital Organization Shimoshizu Hospital Department of Neurosurgery National Hospital Organization Shimoshizu Hospital

Hepatic insufficiency is a fatal liver disease with a significant decrease in functioning hepatocytes. If hepatocytes could be generated from human induced pluripotent stem(hi PS) cells and transplanted into patients with hepatic insufficiency, the disease may become curable. However, a major limitation to this therapeutic strategy is due to the tumorigenicity of hi PS cells and their ability to form cancer. Current methods for eliminating unwanted hi PS cells use genetic manipulation or reagents that are potentially hazardous for hepatocytes; therefore, revised methods are necessary and anticipated. Glucose and arginine are essential cell culture medium ingredients for the survival of most cells, including hi PS cells. However, hepatocytes can produce its own glucose and arginine through galactokinase and ornithine transcarbamylase, respectively. Therefore, it was hypothesized that unwanted hi PS cells could be eliminated in a medium without glucose and arginine, and supplemented with galactose and ornithine instead. This modified medium has been established as hepatocyte selection medium(HSM). So far, attempts to generate a pure colony of mature hepatocytes from hi PS cells have not been successful. After establishment of co-culture in HSM,primary human hepatocytes survive while hi PS cells die within three days. Our latest results regarding a modification of HSM will be introduced in this manuscript.

关键词： Ornithine transcarbamylase Galactokinase Arginine Galactose Urea cycle

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