• Toxicological Research
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• v.9 no.2
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• pp.167-175
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• 1993

Protective effects of dithiol malonate derivatives (DMDs), YH-100, YH-150 and YH-439 on carbon tetrachloride-induced hepatotoxicity were investigated in primary rat hepatocytes culture. Treatment of DMDs to hepatocytes culture did not affect total cytochrome P-450 content and ECOD and AHH activities. Protein and RNA synthesis was also similar to control. Meanwhile, DMDs significantly decreased LDH release and in vitro lipid peroxidation induced by $CCI_4$. Accumulation of cellular triglyceride and decreased secretion of VLDL from liver cells by $CCI_4$ treatment were also significantly protected.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.141-144
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• 2000

Highly open porous polymer matrices are required for high density cell seeding, efficient nutrient, and oxygen supply to the cells cultured in the three dimensional matrices. However, there are severe problems of mass transfer limitations within the cell/scaffolds culture system. Thus we hypothesize that continuos-flow culture conditioning of cells with the scaffolds may improve the cell viability and the differentiated function. In this study, we fabricated porous PLGA scaffolds by using gas-foaming/salt-leaching method as previous described. Viscous PLGA gel paste contains ammonium bicarbonate particulates, acting as a gas-foaming agent as well as a salt-leaching porogen, were cast into Teflon mold and dried. Ammonium bicarbonate salt upon contact to an acidic aqueous solution evloves gaseous ammonia and carbon dioxide by itself. And we conjugated galactose moiety [AGA; $N-(aminobuty1)-O-{\beta}-D-galactopyranosyl-(1{\rightarrow}4)-D-glucoamide]$ to the terminal end group of a PLGA to increase the cell adhesion and matain the differentiated function of hepatocytes. Cell-seeded scaffolds were secured in a flow bioreactor chamber and exposed to continuous flow at 5 ml/min. As a result of our study, the high yield of hepatocytes attachment was accomplished by increasing the concentration of PLGA-AGA conjugate in polymer scaffolds and cells in the scaffolds under continuos flow condition maintained a high level of viability and albumin secretion rate of cultured hepatocytes showed a higher level that of control groups.

• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.4
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• pp.264-268
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• 2001

Aggregates of specific cells are often regarded as better from in artificial organs and mammalian cell bioreactors in terms of cell-specific functionality. In this study, the morphology and liver specific functions of freshly harvested primary rat hepatocytes, which were cultivated as spheroids and entrapped in a synthetic thermo-reversible extracellular matrix, were examined and compared to a control (hepatocytes in single cell form). A copolymer of N-isopropylacrylamide(98 mole % in feed) and acrylic acid (poly (NiPAAm-co-AAC)), a thermo- reversible copolymer gel ma- trix, was used to entrap hepatocytes either in spheroids or single cells. During a 7-day culture pe-riod, the spheroids maintained higher viability and produced albumin and urea at a relatively con-stant rate, while, the single cell culture showed a slight increase in cell numbers and a reduction in albumin secretion Hepatocytes cultrured as spheroids present a potentially useful three-dimensional cell culture system for application in bioartificial liver device.

• Korean Journal of Animal Reproduction
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• v.20 no.3
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• pp.259-269
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• 1996

To establish whether or not androgens is responsible for the induction of vitellogenin(Vg) synthesis and secretion, primary hepatocytes prepared from immature eels were used. The results are follows: 1. Eel hepatocytes were prepared using a collagenase perfusion technique. The isolated cells attached efficiently to fibronectin-coated dishes and subsequently formed monolayers in serum-free medium. These cultures maintained in medium for 10 days with minimal cell loss. 2. Estradiol-17$\beta$(E2) alone was insufficient to induce Vg synthesis. The combination of E2 with methyltestosterone(MT) markedly stimulated Vg synthesis. High vg production occurred in MT concentration from 10-6~10-5M in the presence of E2 (10-6M). Testosterone and androsterone were also effective, but progesterone was not effective in inducing Vg synthesis. Neither MT alone nor testosterone and androsterone alone had any effect on Vg synthesis. 3. E2-primed hepatocytes showed Vg synthesis in both media with and without hormones 1 day after culture. In the cultures with the vehicle, MT, or progesterone, the rate of synthesis seemed to decrease with time. But the combination of E2 and MT showed an intense increase in Vg synthesis. Hepatocytes isolated from E2-primed eels also required androgens for continuating of Vg synthesis. 4. These results demonstrate that androgens act together with E2 in synthesis and secretion of eel Vg.

• Toxicological Research
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• v.7 no.2
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• pp.173-181
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• 1991

Membrane transport of brazilin was investigated i primary cultured rat hepatocytes. Brazilin was transported into hepatocytes very slowly and reached plateau at about 60 minutes. Saturation of transport process was not observed and the transport was not affected by ouabain, metabolic inhibitors, and its structural analog. The amount of brazilin transported into hepatocytes was decreased when the environmental temperature was decreased. These results suggest that brazilin might be transported into hepatocytes by simple diffusional process.

• Animal Bioscience
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• v.36 no.11
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• pp.1693-1699
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• 2023

Objective: Cows that are nursing get around 80% of their glucose from liver gluconeogenesis. Propionate, a significant precursor of liver gluconeogenesis, can regulate the key genes involved in hepatic gluconeogenesis expression, but its precise effects on the activity of enzymes have not yet been fully elucidated. Therefore, the aim of this study was to investigate the effects of propionate on the activity, gene expression, and protein abundance of the key enzymes involved in the gluconeogenesis of dairy cow hepatocytes. Methods: The hepatocytes were cultured and treated with various concentrations of sodium propionate (0, 1.25, 2.50, 3.75, and 5.00 mM) for 12 h. Glucose content in the culture media was determined by an enzymatic coloring method. The activities of gluconeogenesis related enzymes were determined by enzyme linked immunosorbent assay kits, and the levels of gene expression and protein abundance of the enzymes were detected by real-time quantitative polymerase chain reaction and Western blot, respectively. Results: Propionate supplementation considerably increased the amount of glucose in the culture medium compared to the control (p<0.05); while there was no discernible difference among the various treatment concentrations (p>0.05). The activities of cytoplasmic phosphoenolpyruvate carboxylase (PEPCK1), mitochondrial phosphoenolpyruvate carboxylase (PEPCK2), pyruvate carboxylase (PC), and glucose-6-phosphatase (G6PC) were increased with the addition of 2.50 and 3.75 mM propionate; the gene expressions and protein abundances of PEPCK1, PEPCK2, PC, and G6PC were increased by 3.75 mM propionate addition. Conclusion: Propionate encouraged glucose synthesis in bovine hepatocytes, and 3.75 mM propionate directly increased the activities, gene expressions and protein abundances of PC, PEPCK1, PEPCK2, and G6PC in bovine hepatocytes, providing a theoretical basis of propionate-regulating gluconeogenesis in bovine hepatocytes.

• Applied Microscopy
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• v.29 no.4
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• pp.437-450
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• 1999

Bile canaliculi are the structure delivering bile secreted by hepatocytes into the bile passage. Bile secretion is mainly controlled by the cytoskeletal elements, mainly of actin in the microvilli, pericanalicular web. Most studies on the bile secretion have been done in viva situation, however, to control the various parameters in vitro culture system seem to be more useful. To set up an in vitro experimental system, the investigator isolated hepatocytes with an enzymatic method using a mixture of collagenase and hyaluronidase from normal Sprague-Dawley rat liver and cultured. Isolated hepatocytes were round and formed cords in culture. Microvilli covered the whole surface of hepatocytes. Bile canaliculi were formed between hepatocytes and were characterized by the presence of microvilli of various lengths and shapes mainly arising from small surface mounds. Actin filament core in the microvilli and pericanalicular actin web were incomplete. After cytochalasin D treatment, cultured hepatocytes were round but the surface were irregular with surfacen blebs, folds and grooves. Microvilli on the surface were scarce. Bile canaliculi were markedly dilated often with the detached junctional complexes. Bile canaliculi lacks microvilli almost completely and extended into the pericanalirular cytoplasm showing complex vacuolar and tubular structures by transmission electron mciroscopy. Pericanalicular actin web, intermediate filaments were hardly identified. Subsurface actin filaments were scattered scarcely under the cell membranes. These results suggest that hepatocytes isolated from rats can survive and form bile canaliculi in culture and the actin filaments are involved in the formation and/or maintenance of the bile canaliculi.

• Journal of Environmental Science International
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• v.29 no.6
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• pp.633-641
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• 2020

Water temperature is one of the most important factors of fish survival, affecting the habitat, migration route, development, and reproduction. This experiment studied the induction level of heat shock protein (HSP70) mRNA and protein in a walleye pollock (Gadus chalcogrammus) primary hepatocyte culture based on different temperatures. Hepatocytes were attached at 7.5℃ for 24 hours. Hsp70 induction levels were then measured for 48 hours at 5, 8, 11, 14, and 17℃. The induction level was lowest at 5℃ and generally increased with temperature until 14℃. The induction level was reduced at 17℃, indicating that 14℃ is the highest tolerable temperature for hepatocytes. These data indicate that primary hepatocyte cell culture is under no stress at 5 and 8℃. Temperatures greater than 11℃ induce stress, showing similar induction patterns in both mRNA and protein in hepatocytes. The results suggest that 14℃ is the maximum internal defense temperature of walleye pollock survival.

• Biomolecules & Therapeutics
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• v.4 no.3
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• pp.271-274
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• 1996

The antihepatotoxic activity of Bezoar Bovis and Moschus was investigated by in vitro assay method using galactosamine and carbon tetrachloride-induced cytotoxicity in primary-cultured rat hepatocytes. The antihepatotoxic activity was evaluated by measuring the level of glutamate pyruvate transaminase and sorbitol dehydrogenase which were released from the necrotic hepatocytes to the culture medium. In galactosamine-intoxicated hepatocytes, the chloroform fraction of Bezoar Bovis reduced the level of glutamate pyruvate transaminase and sorbitol dehydrogenase resulting in 65% and 59% protection, respectively. The n-Hexane fraction of Moschus resulted in 45% and 40% protection, respectively in this system. In the case of carbon tetrachloride-intoxicated rat hepatocytes, Bezoar Bovis did not have significant effect and only the aqueous fraction of Moschus showed 42% and 40% protection, respectively.

• Korean Journal of Veterinary Research
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• v.50 no.2
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• pp.79-84
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• 2010

The molecule possessing ankyrin-repeats induced by lipopolysaccharide (MAIL) protein is a novel member of the $Ikappa{\beta}$ family. In the present study, we examined the effect of norepinephrine (NE) on MAIL mRNA expression in primary cultured mouse hepatocytes and non-parenchymal liver cells. MAIL mRNA expression in hepatocytes and non-parenchymal liver cells was not directly influenced by NE. However, MAIL mRNA expression in hepatocytes was significantly induced by incubation with a culture medium of non-parenchymal liver cells, treated with NE. Pretreatment with an interleukin (IL)-1 receptor antagonist significantly attenuated the stimulatory effect of the medium. Moreover, exogenous $IL-1{\beta}$ induced MAIL mRNA expression in hepatocytes, while IL-6 and tumor necrosis factor $\alpha$ did not. The concentration of $IL-1{\beta}$ in the medium of non-parenchymal liver cells was significantly increased after NE-treatment. These results suggest that NE can induce MAIL mRNA expression in hepatocytes through $IL-1{\beta}$, released from non-parenchymal liver cells.