Production and release of infectious hepatitis C virus from human liver cell cultures in the three-dimensional radial-flow bioreactor.

Lack of efficient culture systems for hepatitis C virus (HCV) has been a major obstacle in HCV research. Human liver cells grown in a three-dimensional radial-flow bioreactor were successfully infected following inoculation with plasma from an HCV carrier. Subsequent detection of increased HCV RNA suggested viral replication.

Invasiveness of hepatocellular carcinoma cell lines: contribution of membrane-type 1 matrix metalloproteinase.

Intrahepatic metastasis is one of the malignant features of hepatocellular carcinoma (HCC). Matrix metalloproteinases (MMPs) and urokinase-type plasminogen activator (u-PA)/plasmin, are known to be associated with the invasive properties of various types of tumor cells. In this study, we examined which proteinases play a role in the metastatic invasion of human HCC cell lines.

Developments in bioartificial liver research: concepts, performance, and applications.

As an alternative to liver transplantation, numerous researchers have been working toward the goal of development of a fully functional artificial liver. In recent years, artificial liver support systems have been advocated as interim treatments for patients awaiting hepatocyte replacement therapy or liver transplantation; so-called "bridging" treatments. It is recognized that an effective artificial liver system requires: (1) a viable and highly functional hepatocyte cell line, (2) a suitable bioreactor environment and peripheral control systems, and (3) an effective extracorporeal circulatory system to incorporate an artificial liver system.

Involvement of insulin-like growth factor binding protein-3 in the retinoic acid receptor-alpha-mediated inhibition of hepatocellular carcinoma cell proliferation.

We examined the relationship between the expression of retinoic acid receptor-alpha (RAR-alpha) and upregulation of insulin-like growth factor binding protein-3 (IGFBP-3) in the retinoid-induced inhibition of hepatocellular carcinoma (HCC) cell proliferation. HCC cell lines showed a marked expression of RAR-alpha, whereas the expression levels of RAR-beta and RAR-gamma were relatively lower. An RAR-alpha agonist significantly inhibited the HCC cell proliferation both in vitro and in vivo.

Retinol esterification activity contributes to retinol transport in stellate cells.

The mechanisms of retinol transport and accumulation in hepatic stellate cells (HSC) remain to be elucidated. Our previous studies suggested that retinol esterification activity, particularly lecithin:retinol acyltransferase (LRAT) activity, in liver retinoid metabolism is important to elucidate the relationship between retinol uptake by HSC and the esterification of retinol. In the present study, using a human HSC-like cell line, LI90, we demonstrated that retinol esterification activity of LI90 cells is similar to that of primary cultures of rat HSC and higher than that of a human hepatoma cell line.

High density culture of immortalized liver endothelial cells in the radial-flow bioreactor in the development of an artificial liver.

Liver endothelial cells are important components of the tissue along the hepatic sinusoid. They are responsible for microcirculation in the liver and scavenger functions. It would therefore be important to include these cells in any hybrid type of artificial liver in addition to hepatocytes.

Massive culture of human liver cancer cells in a newly developed radial flow bioreactor system: ultrafine structure of functionally enhanced hepatocarcinoma cell lines.

With a view to initiating clinical trials, cell morphology and function for a newly developed artificial liver support system employing highly functional human liver cell line, FLC-7, cultured in a radial flow bioreactor were compared to cells grown in a conventional monolayer culture. The radial flow bioreactor consists of a vertically extended cylindrical matrix comprised of porous glass bead microcarriers through which liquid medium flows from the periphery in toward the central axis generating a beneficial concentration gradient of oxygen and nutrients, while preventing excessive shear stresses or buildup of waste products. The three-dimensional culture system supports high-density (1.

[Work intensity during working hours and different types of care done by special nursing home workers].

This study was done to estimated work intensity during working hours and different types of care to obtain basic data for making a care program. The subjects were care workers in good health (n = 8, 24-45 years) who worked in a special nursing home. The estimated maximal oxygen intake level, which is the maximal aerobic capacity, of each subject was assessed as normal to very good.

Intraocular hemorrhage developing during interferon therapy.

A 43-year-old man with chronic active hepatitis C was treated with interferon-beta (IFN-beta) at a dosage of 6 x 10(6) IU/day for a planned 6-week period. Ocular hyperemia, ophthalmalgia, and increased intraocular pressure in the right eye developed 20 days after the start of treatment. Intraocular pressure remained high, even after discontinuation of IFN therapy, laser therapy, and iridectomy.

[A new liver support system composed of functional human cells and a radial-flow bioreactor].

An artificial liver will be useful for the treatment of acute hepatic failure and a bridge of liver transplantation. The current reports suggest that the hybrid type of artificial liver composed of functional human liver cells and a bioreactor is practical for clinical use. In the present study, we succeeded high density culture on a large-scale of human functional hepatoma (JHH-7) using a newly developed radial flow packed-bed bioreactor.

Regulation of vitamin A transport into cultured stellate cells of rat liver: studies by anchored cell analysis and sorting system.

Stellate cells (SC) in the liver store the most retinoid in the body, but the mechanisms of specific retinoid transport into SC remain to be elucidated. In this study, to analyze the retinoid content of cultured SC, we employed an anchored cell analysis and sorting system (ACAS), which provides fluorescence analysis of single cultured cells under the phase-contrast microscope by utilizing a laser. First, we examined the effect of retinol binding protein (RBP) on retinol transport into cultured SC of rat liver.

Retinol transport in cultured stellate cells of rat liver: studies by light and electron microscope autoradiography.

The mechanisms of specific transport of retinoids into stellate cells (SC) of liver remain to be elucidated. In the present study, we have conducted experiments to observe the intracellular retinoid metabolism of cultured SC of rat liver using light and electron microscope autoradiography (LMARG and EMARG). After 72 h of culture, the cells were incubated in the medium containing 1.

[Consideration on the mechanism of liver regeneration on experimental results].

Liver has enough functional capacity and regeneration ability. Liver parenchymal cells are usually stable, and it is thought being at stage of Go in cell cycle. But liver cells are easily into progressive stage and the liver recovers its functions and volume after partial hepatectomy or liver injury.

Integration of hepatitis B virus DNA into cells of six established human hepatocellular carcinoma cell lines.

The authors successfully established 5 hepatocellular carcinoma cell lines, JHH-1, 2, 4, 5 and 6, derived from hepatitis B surface antigen seronegative patients and one line. JHH-7, from a patient considered to be a hepatitis B virus carrier. In the culture media of any of the JHH cell lines, including JHH-7, hepatitis B surface antigen was not detected by radioimmunoassay.

[Analysis for the integrated hepatitis B virus genome in cells of established human hepatocellular carcinoma cell line JHH-7].

Authors have successfully established 7 human hepatocellular carcinoma (HCC) cell lines. In one of these cell lines, JHH-7, derived from HBs antigen positive HCC, hepatitis B virus (HBV) genome was analyzed by Southern blot hybridization. Digesting with Hind III restriction endonuclease, two bands were obtained at 6.

[Establishment and characterization of a human hepatocellular carcinoma cell line JHH-7 producing alpha -fetoprotein and carcinoembryonic antigen--changes in secretion of AFP and CEA from JHH-7 cells after heat treatment].

A human hepatocellular carcinoma cell line, JHH-7, was established from resected liver tumor of a 53 year old male with hepatitis B virus infection. JHH-7 was composed of polygonal epithelial cells and functionally synthesized and secreted human albumin, AFP, CEA and ferritin. No HBsAg was detected in the culture supernatant of JHH-7 cells.

[Examination of the effects of hyperthermia and anticancer agent on cultured human gall bladder cancer cells by DNA fluorimetric assay].

Tumor cells are generally more sensitive to hyperthermia than normal cells. We applied hyperthermia for bile duct cancer therapy. We also studied the effect of hyperthermia and anticancer agents using cultured cancer cells.

Retinol transport in cultured fat-storing cells of rat liver. Quantitative analysis by anchored cell analysis and sorting system.

It is difficult to study the mechanism of specific transport of vitamin A in fat-storing cells (FSC) in vivo. In this study, transport of vitamin A added to the medium was quantitatively analyzed in cultured FSCs by means of the spontaneous fluorescence emitted by vitamin A. By density-gradient centrifugation with 38% Percoll, an FSC-rich fraction was separated from normal rat liver cells.

Morphological characteristics of human fat-storing cells fractionated and cultured by newly techniques.

Human non-parenchymal cells, especially fat-storing cells (FSCs), were isolated and primarily cultured using 1-2 g of normal human liver tissue obtained in conjunction with tumor biopsies. The human hepatic FSCs were cultured by a modified Howard and Pesch method. Microscopically the cultured human FSCs showed characteristic fat droplets like those in in vivo FSCs.

[Combination therapy of hyperthermia and other methods in liver and bile tract cancers--evaluation of these methods using cancer cell lines in vitro].

In order to evaluate the combination therapy for liver and bile tract cancer, the effects of anticancer drugs and hyperthermia were observed using cultured human cancer cell lines. In the case of gall bladder cancer cell line (NOZ), combination of adriamycin and hyperthermia showed more effective inhibition for cell proliferation than MMC + hyperthermia and 5-FU + hyperthermia. Hepatocellular carcinoma cell line (JHH-4) showed remarkable inhibition of cell growth and secretion of albumin by combination treatment of adriamycin and hyperthermia.

[Protein secretion of human cultured liver cells].

Liver cells have many functions, and one of which is a production of plasma proteins. Therefore, studies on synthesis and production of plasma proteins from hepatocytes are very important for the recognition of various hepatic dysfunctions, clinically. Of late years, a lot of the complex mechanism of protein synthesis and--secretion was elucidated by using a technique of liver cell culture, for example, primary monolayer culture by freshly isolated hepatocytes and cloned cell culture derived from hepatocellular carcinoma.

[Establishment and characterization of human pancreatic adenocarcinoma cell line JHP-1 producing carbohydrate antigen 19-9 and carcinoembryonic antigen].

A new tumor cell line derived from the ascites of a patient with adenocarcinoma of the head of pancreas was established in culture and the nude mouse. The cell line was characterized by the growth with a population doubling time of 22 hr., a high plating efficiency on the plastic surface and a modal chromosome number of 66.

[Effects of TNF on human hepatocellular carcinoma cell lines and their modification by hyperthermia].

Firstly, using HCC cell lines, the effects of r-h TNF were investigated. The authors had already confirmed that these cell lines were derived from human HCC. Each cell line showed a different growth curve on addition of TNF to the culture medium.