• Biotechnology and Bioprocess Engineering:BBE
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• v.3 no.2
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• pp.96-102
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• 1998

Primary hepatocytes of small animals such as rat and rabbit were often used for the study of extracorporeal liver support systems. Freshly isolated rat hepatocytes form spheroids within tow days when cultivated as suspension in spinner vessels. These spheroids showed enhanced liver specific functions and more differentiated morphology compared to hepatocytes cultured as monolayers However, shear stress caused by continuous agitation deteriorated spheroids gradually. In this work we immobilized spheroids to prolong liver specific activities. First, hepatocyte spheroids were suspended in collagen solution containing calcium chloride and then dropped into alginate solution. A thin layer of calcium alginate was formed around the droplet and then was removed after the inner collagen was gelled by treatment of sodium citrate buffer. Spheroids embedded in collagen-gel bead maintained liver specific functions such as albumin secretion rate longer than hepatocyte spheroids exposed to shear stress. Therefore, we suggest that this immobilization technique may offer an effective long-term hepatocyte cultivation and facilitase the development of a bioartificial liver support device.

• Animal cells and systems
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• v.15 no.1
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• pp.19-27
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• 2011

Hepatocytes, the major epithelial cells of the liver, maintain their morphology in culture dishes coated with extracellular matrix (ECM) components such as collagen and fibronectin or biodegradable polymers (e.g. chitosan, gelatin). In these coated dishes, survival of cells and maintaining of liver-specific functions may increase. The aim of this study was to determine a suitable, cost-effective and simple system for hepatocyte isolation and culture which may be useful for various applications such as in vitro toxicology studies, hepatocyte transplantation and bioartificial liver (BAL) systems. In order to obtain primary cultures, hepatocytes were isolated from liver by an enzymatic method and cultured on plates coated with collagen, chitosan or gelatin. Collagen, gelatin-sandwich and gelatin-cell mixture methods were also evaluated. Morphology and attachment of the cells were observed by inverted microscope and scanning electron microscope (SEM). An MTT assay was used to determine cell viability and mitochondrial activity.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.39 no.3
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• pp.303-307
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• 2006

The effects of starvation on histological changes in hepatocytes and the intestinal epithelium in the olive flounder, Paralichthys olivaceus were examined. The starved group was not fed for 12 weeks, whereas the fed group was given food. Food deprivation resulted in a significant decrease in hepatocyte nucleus size and nuclear height in the intestinal epithelium (P<0.05). In the starved group, pronounced degenerative changes were seen in the histology of hepatocytes and the intestinal epithelium; the tissue morphology was irregular and the nuclei were stained basophillic, shrunk irregularly, and very compact. These results suggest that histological changes in hepatocytes and the intestinal epithelium can be used as an indicator of starvation in the olive flounder. In addition, the data are interpreted in detail and some biologically important relationships are discussed.

• KSBB Journal
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• v.7 no.4
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• pp.278-283
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• 1992

Rat hepatocytes were isolated and cultured on the petri dishes treated with various coating materials. Untreated or collagen coated petri dish gave monolayer culture of hepatocyte and proteoglycan, dermatan sulfate, and BSA treated petri dish gave hemispheroid. The untreated Primaria petri dish gave spheroid type of hepatocyte, and heparin and hyaluronic acid treatment gave multilayers. To sustain high cell viability, monolayer cultured hepatocytes was more useful, while it was found that the hemispheroid or spheroid type hepatocytes was more active in the hepatic functions such as ammonia metabolism and albumin synthesis.

• Ocean and Polar Research
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• v.34 no.2
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• pp.165-173
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• 2012

A 26 day experiment was conducted to determine the effects of feeding and starvation on the survival, morphology, and histology in chum salmon (Oncorhynchus keta) fry. We included three experimental groups: starved, fed, and initial. The survival and growth rates were lower in the starved group than in the fed group (P < 0.05). In the starved group, survival began to decline after 16 days, and all fish had died after 26 days. We determined the effects of starvation on the morphometric parameters using the truss and classical dimensions. The dimensions in the head region were larger in the starved group than in the initial and fed groups. In contrast, the truss dimensions of the fed group were larger than those of the initial and starved groups. Starvation reduced the heights of the hepatocyte nuclei and of the intestinal epithelium (P < 0.05). The starved group also showed atrophy of the digestive structures and shrinkage of the foregut and midgut. Starvation led to the degeneration and atrophy of the exocrine pancreas, in which the lumen was markedly diminished and the folds of mucosa were less apparent. The hepatocyte morphology in the starved group was abnormal compared with that of the initial and fed groups, with highly compact, irregularly shrunken nuclei. Melanomacrophages were randomly distributed in the kidneys of the starved group, and their abundance increased rapidly during the experiment. In contrast, neither the initial nor fed group had any melanomacrophages. These results suggest that the nutritional parameters used in this study are useful indices of nutritional status in chum salmon.

• The Journal of Dong Guk Oriental Medicine
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• v.7 no.2
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• pp.97-105
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• 1999

Hepatic tissues of ICR mouse were intraperitoneally injeced with Colpomenia bullosa(CB) Extract after Triton WR-1339(TX) injection were observed to investigate the antihyperlipermic effect of CB extract for hyperlipidemic hepatic tissue caused by destruction of lipid metabolism. The hepatic tissues were obtained at hour-24, 48, and 72 after TX injection with CB extract treatment. And then these specimen were fixed in 10% neutral buffer solution and were cryocut. The tissue stained by H&E for general morphology and sudan black B for lipid distribution. The increase of hepatocyte havinig meshlike cytoplasm were shown in all hepatic lobules after TX injection and the hepatic plates were disappeared in the region of meshlike hepatocyte aggregation. But the hepatocyte having meshlike cytoplasm were disappeared and hapatic plate were rearranged in CB extract injected mouse. The number of blue black colored lipid drop in hepatic cytoplasm of mouse injected with TX were increased and the size of lipid drop were enlarged. But the number of lipid drop in hepatic cytoplasm of mouse treated CB extract were decreased and the size of lipid drop were diminished. As results indicated that the accumulation of lipid drop caused by TX injection were mitigated by the antihyperlipidermic effect of CB extract.

• KSBB Journal
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• v.23 no.5
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• pp.381-386
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• 2008

Lipophilic ammonia is toxic gas and can easily diffuse across cell membranes. Excess ammonia is implicated in the pathogenesis of several metabolic disorders including hepatic encephalopathy and may result in the death. The purpose of this study was to clarify the inhibition effect of metronidazole on liver cell damage due to ammonia in human Hep G2 cell and rat hepatocytes. The effects of metronidazole were studied in ammonium chloride treated human Hep G2 cell (75 mM) and rat hepatocyte (100 mM) following $0.1{\mu}M$ metronidazole treatment. In MTZ+AC group, cell viabilities increased prominently and LDH activities decreased over 25% than AC group. Furthermore, ammonia level according to ammonium chloride treatment reduced over 30% and lipid peroxidation as an index of cell membrane damage decreased more than twice. By comparison with control, catalase activity showed more than 30% reduction in AC group while less than 10% reduction in MTZ+AC group, respectively. In addition, MTZ+AC group showed the similar cell structure as control in cell morphology study by using light microscope, and represented fluorescent intensity decrement compared with AC group in fluorescent microscopic study with avidin-TRITC fluorescent dye. And cleaved PARP expression due to ammonia reduced twofold or more in MTZ+AC group. As the results suggest, metronidazole may protect the liver cell by inhibiting cell damages due to ammonia and be used for an effective antagonist of ammonia in hyperammonemia.

• Toxicological Research
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• v.8 no.2
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• pp.285-302
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• 1992

Human and rat hepatocytes were isolated by nonperfusion method and cultured for longer than 5 days. Human liver biopsy sample and rat liver were used as hepatocyte source. Several physical and chemical factors which were influencing on hepatocyte isolation procedure were examined and a batch isolation procedure was established for small size sample of rat liver. Isolated hepatocytes showed normal morphlologica characteristics in microscopy and electron microscopical examinations and a morphologica response to phalloidin. Isolated cells were cultured as a monolayer and proven to have intact morphological characteristics for longer than 15 days. Because human liver sample is harder and tighter compared with rat liver, a standard procedure for rat hepatocytes was slightly modified to reduce mechanical damage. Similarly with rat hepatocytes, isolated human hepatocytes showed a normal morphological characteristics and could be cultured for longer than 15days. Human and rat hepatocytes were examined on their functional integrities including cytochrome-P450 related enzyme activity and it's inducibility, hormonal inducibility of AIB uptake and TAT activity, albumin synthesis, DNA synthesis, cellular protein maintenance. In all parameters used in the present study, human and rat hepatocytes showed normal functional characteristics.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.5
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• pp.815-820
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• 2004

The effect of capsaicin (0.02%) on the concentrations of liver and serum lipids, GOT and GPT activities, and hepatocyte morphology in male Sprague Dawley rats fed diets with or without erotic acid (1.0%) for 28 days was studied. Liver triglyceride concentration was increased by approximately 4.7-fold after erotic acid-feeding, wherease simultaneous feeding of capsaicin and erotic acid reduced its concentration by 38%. Orotic acidfeeding elevated liver cholesterol concentration, but an addition of capsaicin to the erotic acid diet resulted in the significant decrease in liver cholesterol. Capsaicin alone did not affect the liver triglyceride and cholesterol concentrations. Reduction of the serum concentrations of triglyceride and cholesterol by capsaicin was also observed in rats fed diets with erotic acid. The serum GOT and GPT activities were not significantly different among the experimental groups. Liver tissue morphology showed that the hepatocytes of erotic acid-feeding rats were a typical fatty liver with numerous fat droplets, whereas simultaneous feeding of capsaicin with erotic acid reduced the size and numbers of fat droplets. The present study demonstrated that the diet containing 0.02% capsaicin can prevent the erotic acid-induced fatty liver.

• Applied Microscopy
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• v.28 no.3
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• pp.283-297
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• 1998

Hemopoiesis and morphogenesis of the human fetal liver through from 10 to 32 weeks of gestation were investigated by light and electron microscopy. The results obtained were as follows. Hemopoiesis of fetal liver tissue was found from 10 to 32 weeks of gestation, but the hemopoiesis was decreased at 32 weeks of gestation. At the 32 weeks of gestation, matured erythrocytes were observed in the sinusoid, and formation of liver cell cord and portal triad were established. Differentiation of hepatic cell was characterized by the increase of amount of cell organelles within cytoplasm, decrease of hemopoietic cell, morphological change of nuclear envelope from folding form to round form during the developmental period. These results suggest that human fetal liver plays a hematopoietic function until bone marrow and spleen play their function, but morphology of liver at 32 weeks of gestation was differed with structure observed in liver of adult.