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Biomedical subjects

K Fujise

Publications and source records attributed to K Fujise.

At least 73 records · Page 4Linked to original sources
PubMed1989

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. The FSCs from tumor liver with serious fibrosis contained fewer fat droplets, and the fibrous constituents were especially abundant. The intermediate filaments extending longitudinally were characteristic of the cultured FSCs. In the human FSCs observed by the plasma polymerization replica method, the cells adhered and were stretched relatively thin. Particularly, the ends of the processes adhered and stretched like a folding fan to the bottom of the dish.

Adipose Tissue↗
PubMed1989

[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. Morphologically, JHH-4 cells were enlarged and the nucleus was also enlarged with combination adriamycin and hyperthermia by phase contrast microscopy. Cytoskeleton of JHH-4 cells became irregular and intercellular borderline was unclear by plasma polymerization replica method (PPRM). The effects of BRM (OK-432 and TNF) on HCC cell lines was also investigated. OK-432 directly inhibited proliferation of JHH-4 cells. We observed internalization of OK-432 by JHH-4 cells with TEM and 16-mm movie. TNF showed various effects on human HCC cell lines. Proliferation of two cell lines was inhibited, and one tended to be enhanced after the addition of TNF to the medium. Hyperthermia influenced the effects of TNF to HCC cell lines. We think that this paper is a very significant study for improving the therapy for hepato-biliary cancers.

Antineoplastic Agents↗
PubMed1989

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. The FSCs were observed to retain cytoplasmic fat droplets even on days 3 and 4 of culture. The FSCs containing fat droplets were selected for this experiment by checking their emission of vitamin A fluorescence. To analyze the vitamin A content of isolated cells, we employed a newly developed anchored cell analysis and sorting system (ACAS 470), which provides fluorescence analysis and sorting of adherent cells under the phase contrast microscope by utilizing a laser with its irradiation range narrowed to 1 micron. Vitamin A fluorescence was detectable by this system even in the cultured FSCs. After 24 hours of culture of FSCs in medium with 1 x 10(-6) M vitamin A added, the strength of fluorescence per FSC was 24.3 +/- 11.2 x 10(5)/cell for control, 61.5 +/- 17.6 x 10(5)/cell for retinyl acetate, 26.0 +/- 12.6 x 10(5)/cell for retinyl palmitate, and 59.0 +/- 15.1 x 10(5)/cell for retinol. Thus, retinol and retinyl acetate were transferred to FSCs in significant amounts without the participation of retinol-binding protein. Furthermore, an extended examination was made of the mechanism of the retinol transport observed in this study. Transport was never inhibited by the presence of vitamin E or azide. Retinol may be transferred by passive transport attributable to the concentration gradient rather than by active transport or through cell membrane damage by retinol itself. There was a tendency for inhibition of the transport of retinol into the cells in fetal calf serum. This inhibition may have occurred because the retinol-binding protein or other serum proteins had decreased the concentration of free retinol.

Animals↗
PubMed1989

[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. In this study, we examined the effect of hyperthermia and adriamycin by using newly established cultured human gall bladder cancer cells. In order to calculate cell numbers, we used the new DNA fluorimetric assay with fluorochrome dye Hoechst 33342, developed by Richards et al. For the purpose of confirming the usefulness of this fluorimetric assay, we compared this assay with methylene blue and 3H-thymidine incorporation methods. The following results were obtained: 1) the combination effect of hyperthermia and adriamycin showed an additive effect, 2) the effect of hyperthermia was only intensified according to the extension of heating time, but the combination treatment resulted in a maximum effect at the early stage of heating, 3) DNA fluorimetric assay with Hoechst 33342 dye was a sensitive, easy and rapid method for calculating cell numbers compared with methylene blue and 3H-thymidine incorporation methods, and more useful for studies of cultured cells.

Cell Count↗
PubMed1988

[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. This paper described the results of our observations and other researchers, and then discussed the point of production of human major plasma proteins using the above culture methods, such as albumin, alpha-fetoprotein and transferrin. Furthermore, we showed statistically that half of twenty-six human hepatoma cell lines established until 1988 in Japan, had already lost their secretory potencies of major plasma proteins in vitro.

Carcinoma, Hepatocellular↗
PubMed1988

[Classification of the isolated hepatocytes].

At this present, enzyme perfusion method is a routine technique to isolate hepatocytes from rat liver for the physiological and pathological experiments. This study described a way of the classification of freshly isolated hepatocytes. First of all, the hepatocytes were fractionated with parenchymal and non-parenchymal cells by low speed centrifugation. And then these cells were subfractionated with a newly developed Percoll linear density gradient method. The fractionated parenchymal cells were divided with cells of periportal and centrilobular areas, respectively. Furthermore, their characteristics were confirmed functionally and morphologically. Non-parenchymal cells (NPC) include Kupffer cells, endothelial cells and fat storing cells (FSC, Ito cells). These isolated NPC are fractionated with a method as mentioned above or centrifugal alutriation method. In this paper, fractionation and classification of Kupffer cells and FSC were discussed with the measurement of fluorescent intensity of vitamin A and the morphological observation of cytoskeleton in culture. Especially, transport of vitamin A into FSC were detected autoradiographically.

Animals↗
PubMed1988

[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. JHH-4 exhibited enhancement of growth under the optimum concentration of TNF. On the other hand, growth of JHH-5 and JHH-7 was inhibited by TNF. JHH-7 were more sensitive to TNF than JHH-5, however, the direct effect of TNF on JHH-7 was not potent, as 10(4) u/ml TNF could not prevent proliferation of JHH-7. Morphological examinations were also performed. Phase-contrast microscopy showed that the JHH-4 cells were enlarged and tended to pile up after the addition of TNF to the culture medium. JHH-7 cells became detached from the culture dish due to cell death. Electron microscopy showed irregular proliferation of the rough endoplasmic reticulum of JHH-4 cells and increased number of lysosomes in JHH-7 cells. Furthermore, hyperthermia exhibited an interesting reciprocal action. Proliferation of JHH-4 was inhibited by low concentrations of TNF together with 41.4 degrees C hyperthermia in contrast to the effects of TNF alone. JHH-7 became more sensitive to TNF under hyperthermia at 41.4 degrees C. On the other hand, normal human fibroblast 'HAIN-55' were not affected by TNF at 37.0 degrees C, 41.4 degrees C or 42.5 degrees C. In this paper, the authors tried to study the effects of TNF and hyperthermia on human HCC cell lines.(ABSTRACT TRUNCATED AT 250 WORDS)

Carcinoma, Hepatocellular↗
PubMed1988

[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. The tumorigenicity was proved by the growth in nude mouse and in soft agar. Morphologically the cell line grew as a confluent monolayer with tight adhesion to the plastic surface. Histologically the cell line was epithelial-like in culture and poorly differentiated adenocarcinoma in nude mouse. Ultrastructurally the cell line showed a characteristic pancreatic epithelium. Furthermore, the cell line expressed carbohydrate antigen 19-9 and carcinoembryonic antigen. This cell line, designated JHP-1, has been cultured for at least 100 passages in vitro and maintained for more than 2 years. This cell line would be used as a new model for human pancreatic carcinoma.

Adenocarcinoma↗
PubMed1987

Human bile duct carcinoma cell line producing abundant mucin in vitro.

A human bile duct carcinoma cell line, designated OZ, was established from ascitic effusion of a patient who suffered from obstructive jaundice due to the clogging of the common bile duct with mucinous substances secreted by the cancer cells. OZ was found to be capable of producing mucin in vitro and pools of mucin were macroscopically identified on the monolayer of the cells. On the electron micrographs, cell coat type mucin and abundant intracytoplasmic desmosomes were observed. The OZ cells secreted carcinoembryonic antigen in culture and had high enzymatic activity of gamma-glutamyl transpeptidase. The tumor heterotransplanted into nude mice also showed mucin production.

Adenocarcinoma, Papillary↗
PubMed1987

Severe cardiovascular changes associated with pancuronium after cardiopulmonary resuscitation and after brain injury.

Severe hypertension, tachycardia or ECG changes have been reported following i.v. administration of pancuronium to patients with pheochromocytoma or bronchial asthma. These cardiovascular changes were explained by an interaction between autonomic effects of pancuronium and elevated serum catecholamines or aminophylline. We noted similar cardiovascular changes associated with i.v. administration of pancuronium in two patients after successful cardiopulmonary resuscitation and in two with midbrain hemorrhage and epidural hematoma. In these patients, pancuronium produced no abnormal cardiovascular changes when given during elective surgery or before the occurrence of midbrain hemorrhage. Thus, ischemic brain damage may play a role in producing the severe cardiovascular changes associated with pancuronium.

Aged↗
PubMed1982

Characterization of alpha-fetoprotein secreted from cultured reuber H-35 hepatoma cells.

Reuber H-35 hepatoma cells were examined for their ability to synthesize protein in vitro, especially to produce alpha-fetoprotein (AFP). The presence of AFP in the culture supernatant solution was determined immunologically by the micro-Ouchterlony method. Charge heterogeneity of AFP was examined electrophoretically in continuous gradient polyacrylamide microgels. With regard to the duration of culture, there was no remarkable change in the ratio of two peaks of AFP, and which came out as a major combined peak and a similar peak by PAS staining on the condition of added SDS. These findings indicated that Reuber H-35 hepatoma cells had potential to produce two charge variants of AFP in vitro.

Animals↗