The function of ovarian interstitial cells has been largely addressed using rat theca-interstitial cell culture. However, this preparation is primarily enriched with theca and secondary interstitial cells, which make it difficult to address selectively the function of the primary interstitial cells. We have developed an in vitro culture of hamster ...

Single multifunctional oncoproteins contribute to genomic instability development, but relationships between one or more oncoprotein-associated activities and genetic changes accompanying tumor cell progression are uncertain. Using NIH 3T3 derivative EN/NIH 2-20 containing transcriptionally silent neomycin phosphotransferase gene (neo) integrants with undetectable spontaneous reactivations, we studied wild-type (WT) and mutant adenovirus ...

To establish a continuous cell line, freshly prepared rat parotid acinar cells were stably transfected with a plasmid vector containing the SV40 large T antigen. The acinar origin of these cells was confirmed by Western blotting, enzyme analysis, and morphological analysis. Transformed cells grown in 10% rat serum showed a ...

Animal cells release traces of material onto glass or silicon surfaces during adhesion and migration. This little studied phenomenon is a widespread and normal concomitant of cell migration. The paper introduces the study of such material. The traces can be visualised by different microscopic techniques (e.g. TIRF, IRM, CLSM, AFM, ...

In addition to endothelial cells and vascular smooth muscle cells, we demonstrated that adrenomedullin (AM) is synthesized and secreted from fibroblasts, Swiss 3T3, Hs68, and NHLF cells, in a native and biologically active form. Synthesis and secretion of AM from these fibroblasts was regulated by inflammatory cytokines, such as tumor ...

Peritubular cells, a mixed population of myoid and non-myoid cells obtained by enzymatic treatments of rat seminiferous tubules, are currently utilized as a myoid cell population. We have morphologically and biochemically compared peritubular cells and purified myoid cells isolated from prepubertal rats. The two cell populations appear morphologically similar when ...

Cytotoxic effects of normal mouse serum on mouse tumor cells were investigated in vitro. When FE melanoma cells of C57BL/6 mouse origin, were cultured in medium containing 1% fetal calf serum (FCS) and 10-30% C57BL/6 mouse serum, number of viable FE cells markedly decreased after a little increase in their ...

Several specific cell cycle activities are dependent on cell-substratum adhesion in nontransformed cells, and the ability of the Ras oncoprotein to induce anchorage-independent growth is linked to its ability to abrogate this adhesion requirement. Ras signals via multiple downstream effector proteins, a synergistic combination of which may be required for ...

PC cell derived growth factor (PCDGF) is an 88-kDa glycoprotein purified from the culture medium of the highly tumorigenic mouse teratoma-derived cell line PC. PCDGF was shown to stimulate the proliferation of 3T3 fibroblasts and PC cells. Amino acid sequencing of PCDGF indicated its identity to the precursor for the ...

Metabolism in cultured cells strongly depends on cell density, thus cell density may be an important factor in biotechnological maintenance of cell cultures. Therefore the energy metabolism of three related cell types with different proliferation activity has been characterised: density controlled primary or secondary fibroblasts [pmf], immortalized, but still density ...

Ethanolamine (Etn), as well as its N-methyl (MeEtn) and N,N-dimethyl (Me2Etn) analogues, were recently shown to potentiate the stimulatory effect of insulin on DNA synthesis in serum-starved NIH 3T3 fibroblasts. In the present work we assessed the impact of the co-mitogenic effects of Etn and its methyl analogues on cell ...

Low lethality treatment of the NIH 3T3 mouse cell line with methotrexate (MTX) during exponential multiplication results in heterogeneous, heritable reduction in growth rate of most if not all the replicatively surviving cells. The effective concentrations of MTX are 10 to 100 times higher in molecular, cellular and developmental biology ...

Tissue transglutaminase (tTgase) catalyzes the posttranslational modification of proteins by forming Ca2(+)-dependent intermolecular epsilon (gamma-glutamyl) lysine crosslinks; however, its physiological function is unclear despite increasing evidence for its involvement in the extracellular environment. To define where the enzyme is active and characterizes targets of crosslinking we have modulated tTgase expression ...

A specific antibody combined with a fluorescein-labeled immunoglobulin was used to investigate the topographic distribution of melatonin in a variety of cells of different origins. Positive identification of both nuclear and cytosolic melatonin was confirmed in all the tested cells: Swiss 3T3 mouse fibroblasts, BCG1 bovine granulosa, NB41A3 mouse neuroblastoma, ...

Insulin has pleiotropic effects on the regulation of cellular growth, differentiation, and metabolism. The biochemical events ultimately leading to cell proliferation after insulin treatment have been demonstrated in detail by numerous research groups. However, depending on cell types, it has been shown that insulin has various effects on cell proliferation. ...

The lateral mobility of the neural cell adhesion molecule (NCAM) was examined using fluorescence recovery after photobleaching (FRAP). Various isoforms of human NCAM, differing in their ectodomain, their membrane anchorage mode or in the size of their cytoplasmic domain, were expressed in NIH 3T3 cells and C2C12 muscle cells. When ...

Growth factors and certain oncogenes activate a range of phospholipid-mediated signal transduction pathways resulting in cell proliferation. Demethoxyviridin (DMV), a structural analogue of wortmannin and recently reported as a potent inhibitor of phosphoinositide-3-kinase, inhibited bombesin plus insulin-stimulated increase in cell number in Swiss 3T3 cells, a model of cell proliferation. ...

Rat cholecystokinin-B (CCK-B) receptors were transfected into Chinese hamster ovary (CHO)-K1 (CHO-CCK-B) and Swiss 3T3 (Swiss 3T3-CCK-B) cells, and the effects of receptor activation on cell proliferation and intracellular signaling were investigated. CCK octapeptide (CCK-8) treatment had no effect on cell growth in quiescent CHO-CCK-B cells but inhibited DNA synthesis, ...

Tat is known to act both in a 'cell autonomous' and in 'cell non-autonomous' manner, i.e. Tat can function as a powerful transcription factor both inside the cell in which it is expressed and also in cells not expressing Tat. However, the distance at which Tat can act by the ...

Many lung cancer cell lines are resistant to the growth inhibitory effects of retinoids. However, some small-cell lung cancer cell lines were inhibited by all trans-retinoic acid (ATRA) in serum-free medium. We compared the responses of seven non-small cell lung cancer (NSCLC) cell lines to ATRA in serum-free medium and ...

Retinoylation (retinoic acid acylation) is a post-translation modification of proteins occurring in a variety of mammalian cell lines and in vivo. To gain further knowledge of the role of retinoylation we studied it in NIH 3T3 cells and NIH 3T3 cells transformed by an activated Ha-ras oncogene (NIH Ha-ras-3T3 cells). ...

Murine ecotropic leukemia viruses use a common receptor for entry into host cells; however, the site of virus fusion appears to differ with the host cell. Entry in mouse NIH 3T3 fibroblasts is by endocytosis, whereas entry in rat XC sarcoma cells is by surface fusion. We report here the ...

Mouse cells are not susceptible to infection with echovirus 1 (EV-1) because they lack the viral receptor, human VLA-2. Two mouse fibroblast cell lines, L cells and 3T3 cells, were made susceptible to EV-1 infection after transformation with cDNAs of human VLA-2. After EV-1 infection, L cell transformants of human ...

The effect of a short treatment with keratinocyte-conditioned medium (KCM) on the growth of normal human epidermal keratinocytes was investigated. Serum-free MCDB153 medium was conditioned by keratinocytes for 24 h after plating. Following attachment to uncoated plastic surfaces (4 h after plating), cells were exposed for 20 h to KCM. ...

To investigate whether overexpression of human c-Src leads to cell rounding in anchorage-dependent NIH 3T3 fibroblasts, we have established c-Src-inducible cell lines using a lac repressor-operator system. RN1 cells, which expressed c-Src at a high level after induction, exhibited a spherical morphology and ceased to grow in monolayer culture. RN1 ...

PURPOSE: To examine whether the vitronectin (VN) in serum is responsible for the serum stimulation of phagocytosis in the rod outer segment (ROS) by cultured retinal pigment epithelial (RPE) cells. METHODS: Vitronectin was removed from fetal bovine serum by heparin-agarose affinity chromatography. Concentrations in normal and depleted serum were determined ...

DNA therapeutics show great potential for gene-specific, nontoxic therapy of a wide variety of diseases. The deoxyribose phosphate backbone of DNA has been modified in a number of ways to improve nuclease stability and cell membrane permeability. Recently, a new DNA derivative with an amide backbone instead of a deoxyribose ...

We have examined the importance of cell surface heparan sulfate proteoglycans (HSPG) in fibroblast growth factor (FGF) signaling. 3T3 cells grown under conventional conditions were much more sensitive to FGF-2 compared to FGF-1. However, cells were equally sensitive to FGF-1 and FGF-2 using conditions which reduced the effect of endogenous ...

Hypusine formation on the eukaryotic initiation factor 5A (eIF-5A) precursor is ubiquitously present in eukaryotic cells and archebacteria. In this reaction, deoxyhypusine synthase catalyzes the conversion of one unique lysine residue on eIF-5A to deoxyhypusine using spermidine as the substrate. Hydroxylation of the deoxyhypusine residue completes hypusine formation on eIF-5A. ...

The regulation of protein synthesis was studied in KRC-7 cells (rat hepatoma) grown in complete medium, during serum starvation, and mitogen activation. Upon serum starvation, the cells lost almost completely p67 mRNA, p67 protein, and protein synthesis activity. After phorbol 12-myristate 13-acetate addition, the same serum-starved cells regained p67 mRNA, ...

Sphingosylphosphorylcholine (SPC) has been shown to be a potent mitogen for Swiss 3T3 fibroblasts and also to be an inhibitor of cell growth of some cancer cells, suggesting cell-selective action of the lipid. We examined the effects of SPC, and its structurally-related sphingosine (SP), sphingosine 1-phosphate (S1-P) and membrane-permeable derivatives ...

A cell line of mesodermal origin, FS9, was found to release a Cell Orienting Factor into its culture medium. In contrast with the random migration of controls, the orienting activity causes migrating mesenchymal cells to form an orderly "halo' surrounding tissue explants; individual cells and their cytoskeletons are elongated and ...

The objective of this experiment was to determine the growth characteristics of bovine embryonic muscle cells and to optimize the growth conditions for these cells using commercially-prepared media and sera. In the first study, the growth of muscle cells isolated from the hindlimb was determined by measuring DNA content. The ...

This study showed that nitric oxide (NO)-generating S-nitrosothiols, S-nitroso-N-acetylpenicillamine (SNAP) and S-nitrosoglutathione (GSNO), increased proliferation of BALB/c 3T3 (clone A31-1-1) fibroblasts in vitro. Treatment with SNAP at a relatively low concentration (0.005-0.02 mM) induced an increase in cell number compared to control. SNAP (0.005-0.02 mM) and GSNO (0.025-0.05 mM) both ...

We recently characterized a species of proteochondroitin sulfate (CSPG) secreted by human B cell lines that closely resembles in its structure the serum-derived C1q inhibitor (C1qI). These proteoglycans have in common a molecular mass of approximately 130 to 150 kDa with a core protein of 30 kDa to which up ...

Serum deprivation or exposure of NIH 3T3 cells to actinomycin D (0.25-1.0 microgram/ml; 1 h) was associated with the accumulation of numerous apoptotic cells, as identified by their condensed nuclei and the decrease in cell size. In contrasts, v-H-ras-transformed NIH 3T3 cells were found to be resistant to this apoptosis ...

Incubation of intact U-937 cells with 0.1 microM [gamma-32P]ATP for 10 min resulted in Mg(2+)-independent radiolabelling of about 40 cell surface proteins, some of which became more intensely labelled when cells were differentiated. Presence of 2% newborn calf serum revealed major labelling of three serum proteins with molecular weights of ...

Previous studies demonstrated that inhibition of sphingolipid synthesis by the mycotoxin fumonisin B1 (FB1) disrupts axonal growth in cultured hippocampal neurons (Harel, R., and Futerman, A. H. (1993) J. Biol. Chem. 268, 14476-14481) by affecting the formation or stabilization of axonal branches (Schwarz, A., Rapaport, E., Hirschberg, K., and Futerman, ...

Different established cell lines (SK-v human keratinocytes, B16F10 mouse melanoma, Muntjac fibroblasts, 3T3 mouse fibroblasts, CLV human fibroblasts) as well as primary cultures of mouse fibroblasts and giant cells were treated with N-benzyloxycarbonyl-Ile-Glu(O-t-butyl)-Ala-leucinal (proteasome inhibitor, PSI), a specific inhibitor of the chymotrypsin-like activity of the proteasome. PSI induced well characterized ...

Mobility of anionic residue-containing molecules on 3T3 and simian virus 40-transformed 3T3 (SV40-3T3) cell surfaces was ultrastructurally examined using polycationic ferritin (CF) particles in vitro. CF-binding experiments at 4 degrees C showed continuous distribution of CF particles on entire surfaces of both cells. When 3T3 cells were incubated with CF ...

CD44s (standard isoform) levels and hyaluronan-binding activity were investigated in Balb/c 3T3 cells and their derivatives transformed with ras or sis oncogenes as a function of serum concentration in the medium. 3T3 cells contained low levels of CD44 and did not bind hyaluronan when grown in medium containing 0.5 or ...

The survival of mast cells are dependent on two kinds of growth factors, one derived from T cells (IL-3) and another derived from fibroblasts (stem cell factor [SCF]). The 3T3 fibroblast cell line derived from WCB6F(1-)+/+ mouse embryos (+/+ 3T3 fibroblasts) supported the proliferation of bone marrow-derived cultured mast cells ...

Escape of cancer cells from the circulation (extravasation) is thought to be a major rate-limiting step in metastasis, with few cells being able to extravasate. Furthermore, highly metastatic cells are believed to extravasate more readily than poorly metastatic cells. We assessed in vivo the extravasation ability of highly metastatic ras-transformed ...

An 81 kDa protein was found to be highly expressed in spontaneously immortalized CD1-ICR mouse fibroblasts, RS-4, as compared to normal fibroblasts. RS-4 cells have a reduced serum requirement and exhibit multilayered growth in vitro but are not tumorigenic. The protein was purified from RS-4 cell extracts and used to ...

Previous studies in this laboratory have implicated the membrane transport protein Na/K/Cl cotransporter (NKCC1) as an important component of the signaling pathways activated by phorbol esters in BALB/c 3T3 cells. The NKCC1 protein functions as a Na/K/Cl cotransporter in BALB/c 3T3 cells and many other cell types. Loss of NKCC1 ...

The aims of this study were to investigate in mouse embryonic stem cells (1) the requirement for myo-inositol for cell proliferation, (2) the incorporation of inositol into the phosphoinositides and inositol phosphates of the phosphatidylinositol (PtdIns) signal transduction system and (3) the effect of serum growth factors on PtdIns turnover. ...

Using a model recombinant retrovirus encoding the Escherichia coli lacZ gene, we have found that medium conditioned with NIH 3T3 cells and packaging cell lines derived from NIH 3T3 cells inhibits infection. Most of the inhibitory activity was greater than 100 kDa and was sensitive to chondroitinase ABC digestion, which ...

Sphingosine, a core unit of sphingolipids, has been shown to mediate intracellular signaling events. It is conceivable that sphingosine could act as an intracellular messenger as well as an intracellular modulator, if sphingosine were to be transferred intercellularly. Murine interleukin-2 (IL-2) dependent T-lymphocyte CTLL cells, murine fibroblasts Swiss 3R3 cells, ...

Thrombospondin 1 (TSP1) and thrombospondin 2 (TSP2) are members of the thrombospondin family that have a similar structural organization but somewhat different functional activities. Iodinated recombinant mouse TSP2 bound to NIH 3T3 cells and was internalized and degraded through a chloroquine-inhibitable pathway. TSP2 degradation was saturable, specific, and similar to ...

The rates of cell attachment of the anchorage-dependent mammalian cell line Vero to the gelatin-based macroporous microcarrier Cultispher-G were determined under various conditions. An optimal rate of attachment (0.98 x 10(-2) min(-1)) occurred by an intermittent stirring regimen of 3 min stirring at 40 rpm per 33 min. This stirring ...