• 약학회지
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• 제42권1호
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• pp.108-113
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• 1998

G009, a polysaccharide isolated from the mycelia of Ganoderma lucidum IYO09, showed a hepatoprotective activity against $CCl_4$ induced cytotoxicity in primary cu ltured rat hepatocytes. Incubation of $CCl_4$-intoxicated hepatocytes with G009 significantly reduced the levels of glutamic pyruvic transaminase and sorbitol dehydrogenase released from hepatocytes in the medium. G009 showed antioxidative effect by elevating the activities of glutathione reductase and superoxide dismutase, and the content of glutathione in $CCl_4$-intoxidcated primary cultured rat hepatocytes. Furthermore, G009 significantly elevated glutathione-S-transferase activity in $CCl_4$-intoxicated primary cultured rat hepatocytes. G009 also reduced the production of malondialdehyde, a byproduct of lipid peroxidation. From these results, it could be concluded that G009 exerted hepatoprotective activity against $CCl_4$-induced cytotoxicity through antioxidation.

• Toxicological Research
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• 제3권2호
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• pp.73-80
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• 1987

Primary cultures of adult rat hepatocytes were used to study the cytotoxicity of D-galactosamine. Hepatocytes were isolated by a collagenase perfusion technique and maintained as monolayers in serum-free medium on collagen-coated culture dishes. Treatment of galactosamine to the culture markedly inhibited the uptake of ${\alpha}$-aminoisobutyric acid (AIB) inducible with glucagon and dexamethasone. At0.1 mM of galactosamine, AIB uptake was inhibited significantly when treated for 12 hr. At higher doses (0.25, 0.5 and 1.0mM), a significant inhibition was noticed after 1 hr exposure. Generally the magnitude of the inhibition was related to the dose and treatment time of galactosamine. Treatment of galactosamine also produced a dose- and treatment time-related suppression of the tyrosine aminotransferase (TAT) induction caused by dexamethasone. Meanwhile, uptake of ouabain was not affected by the treatment of galactosamine. The viability of the hepatocytes was decreased only slightly by the treatment of galactosamine; more than 87% of the cells excluded tryphane blue when treated 1 mM galactosamine for 12 hr. Galactosamine induced depressions of AIB uptake and TAT activity were prevented by the simultaneous addition of uridine to the culture. D-Galactosamine, cytotoxicity, hepatocytes culture, ${\alpha}$-aminoisobutyric acid uptake, tyrosine aminotransferase.

• KSBB Journal
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• 제19권3호
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• pp.210-214
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• 2004

본 연구에서는 생분해성 고분자 담체인 PGA 담체에 부착된 간세포의 이식을 통해서 이식된 간세포가 괴사하지 않고 남아 있으며 간 조직 구조의 일종인 담세관 유사구조를 확인하였다. 조직공학적인 간세포 이식 방법의 개발은 간 질환에 새로운 치료방법 개발의 가능성을 열어줄 수 있다.

• KSBB Journal
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• 제7권4호
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• pp.278-283
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• 1992

쥐 간세포를 분리하고 여러가지 물질로 표면이 처리된 용기를 이용하여 배양하였다. 표면처리를 하지 않았을 경우와 collagen으로 처리하였을 경우 간세포는 monolayer형태를 이루며 자랐고, heparin과 hyaluronic acid의 경우에는 multilayer, 그리고 proteoglycan, dermatan sulfate(D. S.), BSA의 경우에는 hemispheroid, 그리고 표면처리를 하지 않은 Primaria는 spheroid형태를 형성하였다. 세포 생존도 면에서는 monolayer가 우수하였고, 간세포의 중요한 대사능이라고 볼 수 있는 암모니아의 제거능이나 albumin생산성 면에서는 hemispheroid 혹은 spheroid가 우수하였다.

• Journal of Microbiology and Biotechnology
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• 제11권3호
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• pp.423-427
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• 2001

Chitosan/alginate capsules were formed by electrostatic interactions and had appropriated mechanical strength, permeability to albumin, and stability to hepatocytes. Rat hepatocytes were isolated and immobilized in chitosan/alginate capsules. During the encapsulation process with hepatocyte, 10% of viability was decreased mainly due to the low pH of the chitosan solution. Among various capsule fabrication methods, the chitosan-alginate capsule showed the highest mechanical strength. Addition of collagen in the capsule with hepatocytes enhanced hepatic metabolism as well as the cell viability for 2 weeks of culture. The hepatocyte in the capsule without collagen decreased the viability to 10% for 2-week cultures.

• Toxicological Research
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• 제2권1호
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• pp.1-7
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• 1986

Procarcinogen induced DNA single-strand breaks and unschduled DNA synthesis were measured in primary rat hepatocytes culture. For DNA single-strand breaks assay, rat liver DNA was prelabeled by injection 3H-thymidine during the peak of DNA synthesis following partial hepatectomy. Hepatocytes were isolated from the rat 2 weeks after surgery by a collagenase perfusion techinique and maintained as monolayers in serum free medium on collagen-coated culture dishes. DNA sigle-strand breaks were measured by the alkaline elution techinique.

• Journal of Microbiology and Biotechnology
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• 제15권1호
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• pp.7-13
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• 2005

Chitosan-alginate capsules were formed by electrostatic interactions and exhibited an appropriate mechanical strength, permeability, and stability for the culture of hepatocytes. Pig hepatocytes were isolated and hepatocyte spheroids formed and immobilized in chitosan-alginate capsules. An encapsulation procedure of 3 min and spheroid formation period of 24 h were the optimum conditions for the best liver functions. Pig hepatocytes with a cell density of $6.0{\tomes}10^6$ cells/ml in the capsules were found to be most suitable for application in a bioartificial liver support system. The encapsulated pig hepatocyte spheroids exhibited stable ammonia removal and urea secretion rates in a bioreactor for 2 weeks. Accordingly, chitosan-alginate encapsulated hepatocyte spheroids in a packed-bed bioreactor would appear to have potential as a bioartificial liver.

• Archives of Pharmacal Research
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• 제18권4호
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• pp.289-292
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• 1995

Icariside II, a flavonol glycoside, was isolated from the aerial part of Epimedium Koreanum Nakai by the anti-hepatotoxic acitivity guided fractionation technique employing $CCl_4-in-toxicated$ primary cultured rat hepatocytes as an assay system. Its anti-hepatotoxic activity was evaluated by measuring activity of glutamic pyruvic transaminase released from the $CCl_4-in-toxicated$ primary cultured rat hapatocytes. Icariside II significantly reduced the activity of glutamic pyruvic transaminase released from the $CCl_4-in-toxicated$ primary cultured rat hepatocytes and resulted in 78% recovery of the toxicity at the concentration of $200{\;}\mu\textrm{m}$. The anti-hepatotoxic activity of icariside II on the $CCl_4-in-toxicated$ primary cultured rat hepatocytes was as potent as that of silybin.

• Animal cells and systems
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• 제15권1호
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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.

• Toxicological Research
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• 제5권1호
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• pp.9-15
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• 1989

The effect of butylated hydroxytoluene (BHT) and its major metabolite, 3, 5-di-tert-butyl-4-hydroxybenzoic acid (BHT-acid) on the uptake of taurocholate into hepatocytes was studied using the primary culture of rat hepatocytes. Hepatocyte were isolated by an in situ collagenase perfusion technique and maintained as a monolayer in serum-free meadia for 24 hours before use. The uptake of taurocholate was saturable with an apparent Km of 12.8+2.8 MuM and Vmax of 0.18+0.01 nmol/mg/min. Both BHT and BHT-acid inhibited the hepatocellular uptake of taurocholate when they were added to the culture.