References

Notes: Several strains have been established from kidney tessues of rhesus monkeys. However, few kidney strains are found originating from cynomolgus monkeys. In the present work, a cell line, designated as the strain JTC-12, was established from kidney tissue of normal cynomolgus female monkey. The culture history, some of the nutritional requirements, the chromosomal constitution and the susceptivility to type 1 poliovirus will be described.
Reference ID: 1713

Reference ID: 2752

Notes: The established cell lines isolated from mammalian kidney were characterized by its receptor activities against hormones and the ability to sythesize sulfolipids localized in the renal tubule. The level of 3':5'-cyclic AMP in JTC-12.P3 (monkey kidney) cells increased in 2 min as much as 2.5-5-jold on activation with 1.0 unit/ml of bovine parathyroid hormone or 1.9 unit/ml of synthetic parathyroid hormone resulting in intracellular cyclic AMP concentration of more than 40pmol/mg protein.
Reference ID: 1691

Notes: A significant difference in the glycosphingolipid composition of JTC-12 P3 cells established from monkey kidney tissue was observed when cells cultured in a protein- and lipid-free synthetic medium containing glucose (DM-160) as a sole carbohydrate source were transferred and cultured in the same medium containing galactose and pyruvic acid (DM-170) in place of glucose. In particular, the amounts of gangliosides GM3, GM2, and GD3 in the cells cultured in DM-170 were 5.3-, 17.8-, and more than 8-fold those in the cells cultured in DM-160, respectively, indicating that anabolism of gangliosides is greatly enhanced in cells cultured in the presence of galactose and pyruvic acid, as compared with cells cultured in the presence of glucose. In fact, after cultivation of cells in the medium with N-acetyl-D- [14C]mannosamine for 96 h, the radioactivity incorporated into the gangliosides of the cells in DM-170 was 10-fold that of the cells in DM-160. Among the gangliosides of the cells in DM-170, highly sialylated molecules such as GD3, GD1a, GD1b, and GT1b were preferentially labeled, indicating that the sialyltransferases responsible for the synthesis of gangliosides are significantly more activated in cells cultured in DM-170 than in DM-160. These observations reveal that the glycosphingolipid composition of the plasma membrane can be modified epigenetically under well-defined conditions and provide important clues for clarifying the roles of glycosphingolipids associated with particular cell functions
Reference ID: 2860

Abstract: Phosphate uptake was studied in confluent monolayers of an epithelial-cell line (JTC-12) derived from monkey kidney. Phosphate uptake consisted of a saturable, Na+-dependent, component, which accounted for about 80% of the uptake, and a nonsaturable, Na+-independent, component. The saturable component was specifically dependent on the presence of extracellular Na+ and has an apparent Km value for phosphate of 0.12 mM at 137-mM-Na+, which is close to those reported in the brush-border membranes in mammalian kidneys. The presence of Na+ in the uptake solution decreased the Km for phosphate without affecting the Vmax. Phosphate uptake was inhibited by carbonyl cyanide p-trifluoromethoxyphenylhydrazone and ouabain, suggesting that phosphate transport is an active, energy-dependent, process and is dependent on an Na+ gradient across cell membranes. With respect to the effect of external Na+ concentration, a sigmoid relation was seen between the initial velocity of phosphate uptake and Na+ concentrations, and Hill analysis gave a Hill coefficient of 1.8. In the pH range 6.6-7.4, phosphate uptake declined with increasing pH. Phosphate uptake was stimulated when cells were cultured in the presence of insulin, and was also affected by changes in phosphate concentrations in cultured medium. These results indicate that JTC-12 cells have an Na+-dependent phosphate-transport system with many of the features of phosphate transport in the proximal tubule
Reference ID: 5530

Abstract: The JTC-12 cell, an established cell line derived from a normal monkey kidney, was studied in an attempt to characterize the epithelial qualities. Phase contrast microscopy showed dome formation in confluent monolayers and electron microscopic examinations revealed the presence of numerous microvilli on the apical membranes and desmosome between cells. Sonicated cells showed activities of gamma-glutamyl transpeptidase, leucine aminopeptidase, alkaline phosphatase, and trehalase, marker enzymes of renal proximal epithelium. Alkaline phosphatase activity exhibited the characteristics of a renal type isozyme. Furthermore, confluent JTC-12 monolayers exhibited Na+-dependent transport of hexose, amino acid as well as inorganic phosphate. These findings indicate that JTC-12 cells in monolayer culture maintain ultrastructural, biochemical, and physiological properties of renal proximal epithelial cells. This cell line will be useful for further studies on cellular functions of renal proximal epithelium
Reference ID: 5531

Abstract: Cultured monkey kidney cells (JTC-12) have a Na+-dependent phosphate (Pi) transport system with characteristics similar to that of the renal proximal tubule. Na+-dependent Pi uptake in JTC-12 cells is affected by Pi concentrations in the culture medium. In this investigation, further characterization of this phenomenon was carried out. Lowering the concentration of extracellular Pi (3.0 mmol/l to 0.3 mmol/l) induced an increase in Na+-dependent Pi uptake compared with that in control cells maintained in 3.0 mmol/l Pi, whereas Na+-dependent transport of hexose and amino acid was not altered. This response was first evident at 4 h after the extracellular Pi concentration was reduced and slowly developed over the subsequent 24 h. Kinetic analysis showed an increase in the Vmax without a change in the apparent Km for Pi in cells cultured in the low Pi concentration compared with control cells. The response of Pi uptake was only partially prevented by cycloheximide, suggesting that both protein synthesis-dependent and -independent mechanisms are involved in the development of the response. Insulin, which has a stimulatory effect on Pi uptake in JTC-12 cells, did not affect this response. These data indicate that JTC-12 cells respond to changes in extracellular Pi concentration by changing the Na+-dependent Pi uptake system. This response has a number of properties typical of the phenomenon of adaptation of renal Pi transport in vivo to dietary phosphorus load
Reference ID: 5532

Notes: Third Department of Internal Medicine, Mie University School of Medicine, JapanFAU - Baba, M
Abstract: Human hepatitis A virus (HAV) derived from 10% HAV infected marmoset liver homogenate and faeces from acute hepatitis A was successfully propagated in vitro in a new cell line, JTC-12.P3. The cell line originated from the renal cortex of cynomolgus monkey which was adapted to growth in a serum free, protein free, chemically defined synthetic medium. Replication of the virus was followed by solid phase RIA, immunofluorescent staining, and immunoelectron microscopy. The propagation of HAV occurred over several passages, with the 1st and 2nd passages requiring at least 8 weeks each. However, with the increasing serial passage of virus, the period needed to detect it was shortened, suggesting the adaptation of HAV to the cells. The identity of the newly synthetized virus particles with HAV was established by immunoelectron microscopy and immunofluorescent blocking effect with human convalescent serum. The HAV propagated in JTC-12.P3 cells banded predominantly at a density of 1.32 g/cm3 in CsC1 gradient. The infected cells showed no specific signs of CPE. Ultrastructurally, clusters of virus particles 27 nm in diameter were observed mainly in the lysosomal vesicles and freely in crystalline array in the cytoplasm, too. Addition of 0.1% of various anti-HAV negative sera or of prostaglandin E1 to the culture medium caused accelerated propagation of HAV
Reference ID: 5533

Abstract: The induction of alkaline phosphatase (ALP) by dibutyryl adenosine 3':5'-cyclic monophosphate (Bt2cAMP) was investigated in strain JTC-12 . P3 cells derived from monkey (Maccaca irus) kidney cortex. ALP activity was increased by Bt2cAMP in a dose-dependent manner, reaching a plateau at concentrations higher than 5 mM with the activity being about 4 times that of the controls. The concentration of Bt2cAMP required for half-maximal induction of ALP activity was about 0.8 mM. ALP activity was increased rapidly by Bt2cAMP for the first 5 days and then continued to increase gradually towards a plateau level. Removal of Bt2cAMP from the medium caused a rapid decrease in the activity, suggesting that the induction of ALP activity by Bt2cAMP is reversible. ALP activity was induced synergistically in the presence of 1 mM sodium butyrate together with Bt2cAMP at concentrations from 0.01 to 1 mM. It was also found that in the presence of 1 mM Bt2cAMP, sodium butyrate increased ALP activity in the same manner as Bt2cAMP did in the presence of 1 mM sodium butyrate. Although dexamethasone, a potent glucocorticoid, had no effect on ALP activity in control cells, the hormone suppressed the ALP activity induced by Bt2cAMP in a dose-dependent manner. At concentrations above 0.2 mM, two xanthine derivatives, theophylline and 3-isobutyl-1-methyl-xanthine (IBMX), also inhibited the induction of ALP activity by 1 mM Bt2cAMP. Inhibitors of protein synthesis, cycloheximide (1.5 micrograms/ml) and pactamycin (10 micrograms/ml), as well as inhibitors of RNA synthesis, actinomycin D (2 micrograms/ml) and alpha-amanitin (50 micrograms/ml), suppressed the induction of ALP activity
Reference ID: 5534