The replica filter technique has been used to isolate variants of Swiss mouse 3T3 cells which produce heparan sulfates with altered levels of sulfation. These changes in the extent of sulfation correlate with alterations in cell morphology, in the organization of cytoskeletal elements, focal contacts, and the extracellular matrix, and ...

Using benzo(a)pyrene (BaP) as a probe for aryl hydrocarbon hydroxylase (AHH) activity, differences in mixed-function oxidase (MFO) activity were observed using microspectrofluorimetry in single living cells during long term treatment with 3-methylcholanthrene (3-MC) or carbaryl. Although these two compounds differ in chemical structure, similar effects were observed in 3T3 cell ...

There is growing evidence that cell shape regulates both proliferation and differentiated gene expression in a variety of cell types. We have explored the relationship between the morphology of articular chondrocytes in culture and the amount and type of proteoglycan they synthesize, using cytochalasin D to induce reversible cell rounding. ...

The interaction of cells with solid surfaces is important in many settings, including the response of tissue to implanted materials. Protein adsorption to the surface plays a critical role in controlling cell interactions with surfaces. However, few comprehensive studies of both cell behavior and protein adsorption in complex protein mixtures ...

The growth rate of spontaneously transformed BALB/3T3 cells is proportional to glutamine concentration between 50 and 400 microM, with little or no growth occurring in less than 50 microM glutamine. By contrast, nontransformed BALB/3T3 cells multiply, although slowly, with as little as 20 microM glutamine. Neither cell type depletes the ...

When highly-purified nuclei of Swiss-mouse 3T3 cells are incubated with [32P]-gamma ATP, radioactivity is incorporated into phosphatidic acid and the two polyphosphoinositol lipids, phosphatidylinositol(4)P and phosphatidylinositol(4,5)P2. If the cells are pre-treated with insulin-like growth factor I, this incorporation into the phospholipids is decreased. The effect is maximal by 2 minutes, ...

Malignantly transformed mouse fibroblasts synthesize and secrete large amounts of major excreted protein (MEP), a 39,000-dalton precursor to an acid protease (cathepsin L). To evaluate the possible role of this protease in the transformed phenotype, we transfected cloned genes for mouse or human MEP into mouse NIH 3T3 cells with ...

When separated and proliferating rat 3T3 cells are treated with butyrate (6 mM), DNA synthesis stops within 24 h, while RNA and protein synthesis proceed unaffected. This gradually converts normal cells into giant ones in the presence of butyrate (volume up to 30-fold greater). The giant cells stop growing when ...

A culture of BALB/c 3T3 cells grown in the presence of 40 microM of the ceramide analog compound RV538 (2-decanolyamino-3-morpholino-1-phenylpropanol) for several passages caused a substantial decrease in the level of all glycosphingolipids and an accumulation of ceramide and sphingomyelin. Associated with these chemical changes of sphingolipid composition and metabolism, ...

Cultures of rat astroglial (AG) cells treated with the ganglioside GM1 in serum-free medium respond with an increase in DNA labeling and cell proliferation. However, GM1 doses above 60 microM cause decreasing DNA labeling to levels even below that determined in the absence of added GM1. Quantitative determination of cell ...

Human (D54Mg, GaMg) and rat (BT5C, BT4Cn) glioma cells cultured in a chemically defined medium showed reduced growth when compared to serum-supplemented medium. The BT5C cells changed from a flat epithelioid morphology to a more glia-like structure. The serum-free medium caused an aggregation of BT5C cells which spontaneously lost anchorage ...

Cell quiescence resulting from culture of normal human fibroblasts in low serum (0.5%) was associated with a subsequent gradual increase in the expression of the cell-surface glycoprotein VLA-1, and a corresponding decrease in the expression of extracellular matrix adhesion receptors VLA-2 and VLA-3. Quantitation using either flow cytometry or immunoprecipitation ...

To investigate whether resting cells of 3T3 mouse fibroblasts carry out de novo synthesis of deoxyribonucleoside triphosphates, we determined the turnover of the thymidine triphosphate pool of G0 cells obtained by starvation of cultures for platelet-derived growth factor. These cells were contaminated by less than 1% S-phase cells. In the ...

The intracellular level of DNA topoisomerase II appears to be reversibly regulated by serum concentration in cultured primary human skin fibroblasts (HSF). Upon serum starvation, the intracellular level of topoisomerase II in HSF, as monitored by immunoblotting with antitopoisomerase II antibodies, gradually decreased to a nondetectable level (less than 10(4) ...

Data presented here demonstrate that recombinant gamma interferon (rIFN-gamma) activated a single population of 10% fetal calf serum-elicited mouse peritoneal exudate cells to express tumoricidal activity but not bactericidal activity for the facultative intracellular bacterium Listeria monocytogenes. Fetal calf serum-elicited cells incubated with rIFN-gamma phagocytosed listeriae normally, suggesting that their ...

Mitogen (PHA)-induced proliferation of peripheral blood mononuclear cells (PBMC) was reduced by more than 70% 2 h after the haemorrhage of 30% of blood volume. Experiments using isolated macrophages and lymphocytes showed that post-haemorrhage macrophages were functionally normal and that lymphocytes were responsible for the observed haemorrhage-induced depression of proliferative ...

We have previously shown that normal chicken serum (NCS) is able to interfere with the IL 2 promoted incorporation of DNA precursors into T lymphoblasts and that serum derived from autoimmunity prone Obese strain (OS) chickens is deficient in this respect. This "defect in non-specific suppression" has been speculated to ...

Experiments on the introduction of the regulatory subunit of cAMP-dependent protein kinase type II (RII) into NIH 3T3 cells clearly demonstrated its translocation into the nucleus. The labelled protein was incorporated into erythrocyte ghosts and their fusion with the cells was carried out. The dynamics of distribution of the labelled ...

Acidic fibroblast growth factor (aFGF), a polypeptide with a mol. wt of approximately 16,000, is a potent mitogen for a variety of cells and shares 55% amino acid sequence identity with basic FGF. The recent isolation of three new oncogenes which share 35-45% amino acid sequence similarity with the FGFs ...

The rate of attachment of chick embryo fibroblasts (CEF) incubated for 1 h at pH 6 in the absence of serum, was equal to about 90% with 2% of spreading cells. The addition of 1% horse serum (HS) decreased to about 30% the rate of attached cells but increased to ...

Bradykinin-stimulated prostaglandin synthesis was investigated in Swiss albino 3T3 fibroblasts (Swiss 3T3 cells) and bovine pulmonary artery endothelial cells (CPAE). Previous studies have indicated that bradykinin stimulates arachidonic acid release in Swiss 3T3 cells by activating phospholipase A2 and by activating phosphatidylcholine-specific phospholipase C in CPAE cells. The dose-response for ...

The effects of cell density and growth in 10% foetal bovine serum and 10% newborn calf serum on the activity of the enzyme (Na+ + K+)-ATPase were studied in 3T3 and SV3T3 cells. The enzyme activity decreases in 3T3 cells grown in foetal bovine serum as the cells approach confluency ...

We have detected and partially characterized factors that promote the directed migration of mouse epidermal cells in a modified Boyden chamber assay. Smooth muscle cells grown in culture were found to secrete a potent chemoattractant for epidermal cells. This activity was further characterized and compared to the chemotactic activities found ...

At low serum concentrations (3% or less), individual Swiss 3T3 cells display marked heterogeneity in proliferative capacity. Here we show that this heterogeneity arises at extremely high frequency within a clone, often with sister cells showing considerable differences in capacity for further proliferation. The heterogeneity is unlikely to be due ...

The turnover of phospholipids was investigated in quiescent serum-starved Chinese-hamster ovary (CHO-K1) cells stimulated to progress through the cell cycle by the addition of dialysed bovine serum. A variety of radiolabelling techniques were employed to study the rapid effects of serum on phospholipids and later events during G1 and S ...

A method is described for the determination of cellular F-actin content using fluorometry of TRITC-phalloidin at very high dilution. In this case no saturation of all binding sites available is reached, however the staining is highly specific and the specificity is not affected by the preparative procedure as may be ...

A highly active adipogenic agent was identified in an in vitro adipose conversion system. This agent, ADD 4743 (or ADD), was synthesized by Takeda Chemical Industries (Osaka) as a 3-hydroxy derivative of an oral antidiabetic agent, ciglitazone, and has been presumed to be an active metabolite of the latter substance. ...

To evaluate the proteolytic activities of oncogene-transfected 3T3 cells, we have developed a copolymerized substrate electrophoretic assay that permits the detection of picogram quantities of proteases produced by cells in culture. Our assay involves a gelatin substrate copolymerized in a polyacrylamide gel. Purified cell membrane preparations were run on gels ...

Alteration of the growth properties of the established murine fibroblast cell lines NIH 3T3 and 3Y1 was studied in monolayer cultures and in cells suspended in semisolid medium after introduction of a cloned human papillomavirus type 16 (HPV16) DNA. HPV 16 DNA stimulated both cell lines to grow beyond their ...

15-Hydroxyeicosatetraenoic acid (15-HETE), a major lipoxygenase metabolite of arachidonic acid in fetal bovine aortic endothelial cells, was a mitogen for these cells, stimulating both cell proliferation and DNA synthesis in the presence of serum and serum-deprived cells. In [14C]arachidonic acid-labeled confluent endothelial cell monolayers, 15-HETE (30 microM) caused an elevation ...

We have further characterized a recently described B-cell stimulatory factor that contains chondroitin sulfate proteoglycan and 70- to 75-kDa protein, both of which are secreted by T cells and coisolate (T-cell proteoglycan fraction, T-PGF). Using T-PGF isolated from a T-cell hybridoma (T14), it was observed that the association between B-cell ...

The effect of mitomycin C on the accumulation of specific mRNAs was studied in asynchronously growing Swiss 3T3 cells, as well as in synchronously growing serum stimulated ts13 cells (a temperature-sensitive mutant from the BHK cell line). It was observed that the steady-state level of p53 RNA experienced some increase ...

The level of gap-junctional (cell-cell) communication was studied by the radioisotope transfer technique in NIH 3T3 cells exposed to NiSO4, MgSO4, or both salts combined. Monolayered NIH 3T3 donor cells were labeled with [3H]-uridine for 3 h and then co-cultured with non-labeled recipient NIH 3T3 cells for 3 h in ...

Insect cell lines are not well-suited to colony formation in liquid medium following low-density cell plating. The present studies demonstrate that the time of addition of fetal bovine serum to the culture medium and the number of gamma-irradiated feeder cells added to each plate are important factors in developing a ...

When the glycosaminoglycans (GAGs) of BALB/c 3T3 and B16.F10 melanoma cells were compared during parallel studies, substantial differences were found in the cellular content, rates of turnover, and extent of intrinsic sulfated GAG degradation between the two cell types. The 3T3 cells had more than 10-fold more metabolically-labeled [35S]-GAGs per ...

A factor specifically inhibiting the hepatocyte cell cycle in vivo was found to block the G1-S transition of liver cells in vitro. It proved to be non-toxic in our culture conditions, as judged by the reversibility of the effect on cell proliferation. It was not active on DNA synthesis in ...

Proliferating 3T3 mouse fibroblasts contain higher levels of the lectin carbohydrate-binding protein 35 (CBP35) than do quiescent cultures of the same cells. An immunofluorescence study was carried out with a rabbit antiserum directed against CBP35 to map the cellular fluorescence distribution in a large population of cells under different growth ...

The bombesin receptor present on the surface of murine and human cells was identified using 125I-labeled gastrin-releasing peptide as a probe, the cross-linking agent disuccinimidyl suberate, and sodium dodecyl sulfate gels. A clone of NIH-3T3 cells which possesses approximately 80,000 bombesin receptors/cell with a single binding constant of approximately 1.9 ...

Most in vitro T cell proliferation experiments are performed by using serum-supplemented medium, yet the actual contributions of serum components to cell cycle progression remain ill-defined, thus complicating attempts to fully define requirements for cell division. By utilizing a functional separation between T cell receptor-triggered "competence" and IL 2-promoted "progression" ...

Tumour-promoting phorbol esters and diacylglycerols have a variety of biochemical and biological effects in common in a number of cell culture systems; thus, it has been suggested that diacylglycerols are endogenous functional analogues of phorbol esters. In order to examine more directly the possible relevance of these functional analogues in ...

When highly enriched multipotential spleen colony forming cells (CFU-S) obtained following fluorescence activated cell sorting (FACS-CFU-S) are cultured on marrow stromal cells, they undergo proliferation and development to produce mature haemopoietic cells (Spooncer et al., Nature, 316:62-64, 1985). We now show that FACS-CFU-S behave in a similar way when cultured ...

Quiescent serum-starved 3T3 cells can be stimulated to initiate DNA synthesis after addition of conditioned media from spontaneously tumor-transformed 3T3 cells (3T6-cells) or from SV-40-transformed 3T3 cells (SV-3T3 cells). The conditioned media were found to stimulate both the chromosome cycle (i.e., DNA synthesis and cell division) and the growth cycle ...

The binding of [125I]gastrin releasing peptide ([125I]GRP) to Swiss 3T3 cells at 37 degrees C increases rapidly, reaching a maximum after 30 min and decreasing afterwards. The decrease in cell-associated radioactivity at this temperature is accompanied by extensive degradation of the labelled peptide. At 4 degrees C equilibrium binding is ...

A method is described for the analysis of cell shape, using an image analyzer connected to a computer to assess the cell outline. A series of parameters to assess the contribution of large cytoplasmic expansions to cell morphology and to cell spreading on a planar substratum were used to quantify ...

The relationship between growth, protein degradation, and cellular autophagy was tested in growing and in growth-inhibited 3T3 cell monolayers. For the biochemical evaluation of DNA and protein metabolism, growth-inhibited 3T3 cell monolayers with high cell density and growing 3T3 cell monolayers with low cell density were labeled simultaneously with [14C]thymidine ...

Techniques to determine intracellular pH generally report the average pH of population and do not indicate whether or not there is significant variance among cells within the population. Population variance is important to ascribe pH changes on a per cell basis. The magnitude of the pH change in individual cells ...

LLC-PK1 cells can be brought into a classical quiescent state by depriving them of serum for 6 days. At this time, pulse-labeling with [3H]-thymidine shows that only 3% of the cells are synthesizing DNA, but the quiescent cells can be stimulated with serum to re-enter the cell cycle at a ...

We studied the interactions between the perfluorinated fatty acid nonadecafluoro-n-decanoic acid (NDFDA) and the cell wall-less procaryote Acholeplasma laidlawii, which were cultured in an identical medium base but with different serum supplements. When grown in mycoplasma media supplemented with PPLO serum fraction (Difco Laboratories, Detroit, Mich.), A. laidlawii was rapidly ...

Liposomes containing phosphatidic acid were capable of stimulating DNA synthesis in quiescent Swiss 3T3 cells while liposomes composed of other phospholipids were not. These results show that liposomes, which are usually employed to deliver non-lipid molecules into cells, can themselves have profound effects on cell growth. The possible mechanism of ...

Quiescent mouse NIH 3T3 cells responded to microinjection of activated ras p21 with a rapid and sustained rise in intracellular pH (approximately 0.17 pH units). The p21-induced pH change was inhibited by amiloride treatment or growth of cells in medium low in sodium, suggesting a role for the Na+/H+ antiporter. ...