Physiological quiescence in plasma‐derived serum: Influence of platelet‐derived growth factor on cell growth in culture

Ross, Russell; Nist, Cynthia; Kariya, Beverly; Rivest, Mary Jane; Raines, Elaine W.; Callis, Jim

doi:10.1002/jcp.1040970325

Cited by 128 publications

(75 citation statements)

References 20 publications

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“…2) . Synthetic state cells maintained quiescent in 5% PDS for 24 d and then challenged with 5% WBS respond within 36-48 h by logarithmic growth (27) .…”

Section: Spontaneous Change In Phenotypementioning

confidence: 99%

“…They do not contract in response to angiotensin II or norepinephrine or to electrical or mechanical stimulation (7,17,18) but readily synthesize collagen, elastin, and glycosaminoglycans (1,2,16,22,23,32) and are thus in a synthetic state . Subcultured aortic smooth muscle cells exposed to the platelet-derived growth factor (PDGF) in whole blood serum (WBS) proliferate in a dose-dependent manner with cell division commencing within 24 to 36 h of challenge (25)(26)(27)(28), and smooth muscle which has migrated from aortic explants, proliferated, and reached a stationary growth phase can be stimulated to another proliferative phase by increased amounts of PDGF, hyperlipemic serum, or its low-density lipoprotein (LDL) within 1-2 d of challenge (8)(9)(10)(11)) .…”

mentioning

confidence: 99%

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Phenotype-dependent response of cultured aortic smooth muscle to serum mitogens.

Chamley-Campbell¹,

Campbell²,

Ross³

1981

The Journal of Cell Biology

426

158

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Smooth muscle cells from the aortic media of adult pigs and monkeys have been grown in primary culture by plating cells enzymatically dissociated from the intact aorta. During the first 6 d these cells are in the "contractile" phenotype. That is, they contract slowly in response to angiotensin II and their cytoplasm is filled with both thick and thin myofilaments . In this state they do not incorporate [3H]thymidine into DNA or proliferate in response to normolipemic or hyperlipemic whole blood serum (WBS) . After 7 d in culture the cells undergo a spontaneous modulation of phenotype to a "synthetic" state where they cannot be stimulated to contract and their cytoplasm is filled with organelles usually associated with synthesis of secretory protein . Thick myosin-containing filaments can no longer be demonstrated . When challenged with normolipemic or hyperlipemic WBS the cells incorporate [3H]thymidine into DNA and undergo logarithmic growth . It is suggested that when smooth muscle is in the contractile phenotype (as normally exists for most cells in the aortic media of adult animals) it does not divide when challenged with serum mitogens but can undergo a change of phenotype to a synthetic state in which division can be stimulated .Differentiation of a cell involves transition from an initially multipotential state to the specialized form typical of the adult . A fully differentiated cell is one therefore that has lost all potentiality to develop into another cell type . The concept of cell modulation encompasses the fact that a fully differentiated cell can assume a different function (with associated morphological changes) in response to an altered environment without any change in its type-specific character. Modulation is therefore reversible.The smooth muscle cell is the only cell type present in the media of mammalian arteries (19). It must therefore be responsible not only for maintaining artery wall tension via contraction-relaxation but also for vascular remodeling, repair, and growth . These latter functions require the retention in the adult of certain basic mesenchymal properties, namely the abilities to synthesize extracellular matrix and to divide (33). This multiplicity of functions requires a whole spectrum of variation in morphology or phenotype. At one end of the spectrum is the smooth muscle cell whose major function is contraction . The cytoplasm of this cell is largely filled with thick and thin myofilaments and it has been described as being in the "contractile" state (6) . The vast majority of cells in the media of the adult aorta are in this phenotype. At the other end of the spectrum is the smooth muscle cell engaged in the synthesis of extracellular matrix and/or division. The cytoplasm of this cell

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“…2) . Synthetic state cells maintained quiescent in 5% PDS for 24 d and then challenged with 5% WBS respond within 36-48 h by logarithmic growth (27) .…”

Section: Spontaneous Change In Phenotypementioning

confidence: 99%

mentioning

confidence: 99%

Phenotype-dependent response of cultured aortic smooth muscle to serum mitogens.

Chamley-Campbell¹,

Campbell²,

Ross³

1981

The Journal of Cell Biology

426

158

View full text Add to dashboard Cite

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“…In addition, the Ross group regularly (25)(26)(27) (31) reported appreciable proliferation of 3T3 cells in the presence of low concentrations of nondialyzed plasma and Gospodarowicz et al (32) reported considerable proliferation in the presence of physiological (5-10%) concentrations of dialyzed plasma.…”

mentioning

confidence: 99%

Active proliferation of Rous sarcoma virus-infected, but not normal, chicken heart mesenchymal cells in culture medium of physiological composition.

Balk¹

1980

Proc. Natl. Acad. Sci. U.S.A.

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Normal as well as Rous sarcoma virus-infected chicken pectoral and chicken embryo fibroblasts proliferate actively in a plasma-containing medium of physiological ion concentrations (Ca2+, 1.2 mM; Mg2+, 0.7 mM). Reduction of medium calcium and magnesium concentrations is necessary to achieve selective quiescence of normal fibroblasts in these cell systems. By contrast, normal chicken heart mesenchymal cells proliferate only sluggishly (one doubling or less during a 6-day period) in a plasma-containing medium of ph siologic ion concentrations, whereas Rous sarcoma virus-infected heart mesenchymal cells proliferate actively (more than four doublings during an initial 2-day phase of exponential growth). The chicken heart mesenchymal cell system therefore has great potential for studies of the mechanism that initiates cell replication and of the failure in cellular regulator processes that is responsible for the autonomous initiation of replication of neoplastic cells. From comparison of the chicken heart mesenchymal cell system to dialyzed plasma-based systems in which 3T3 cells tend to proliferative quiescence, it is argued that this proliferative quiescence of 3T3 cells is a result of cell starvation and is not physiologically meaningful. It has been demonstrated (1-3) that, in a plasma-containing culture medium of reduced calcium and magnesium concentrations, normal chicken pectoral muscle fibroblasts are proliferatively quiescent whereas Rous sarcoma virus (RSV)-infected pectoral fibroblasts proliferate actively. In medium of physiological ion concentrations, both normal and neoplastic pectoral fibroblasts are proliferatively active. On the basis of these findings, we hypothesized (1-3) that cytosolic Ca2+ and Mg2+ are involved in the initiation of cell replication and that failure of cytosolic divalent cation homeostasis contributes to the autonomous initiation of replication of neoplastic cells.In the course of studying the proliferative behavior of mesenchymal cells cultured from tissues other than pectoral muscle, it was found that RSV-infected chicken heart mesenchymal cells proliferate actively but normal chicken heart mesenchymal cells do not in a plasma-containing culture medium of physiologic ion concentrations. This provides a culture system in which, under physiological conditions, normal cells are essentially quiescent but their rigorously comparable, stably neoplastic counterparts proliferate actively. MATERIALS AND METHODSExperiments were performed on secondary cultures of normal and RSV-infected chicken heart mesenchymal cells, chicken pectoral muscle fibroblasts (1-3), and chicken embryo fibroblasts. The general materials and methods have been described (1-4). The culture medium (4) was modified by inclusion of folinic acid at 0.02 mg/liter, taurine at 8 mg/liter, and carnitine at 10 mg/liter; this modified medium (Table 1) contains all of the major plasma crystalloids at physiological concentrations. Unless otherwise stated, cultures were in synthetic medium plus heparin-treated plasma (...

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“…Cells were grown to Ͼ90% confluency and medium was changed every 4 days. Cell synchronization was achieved by serum deprivation (0.5% fetal bovine serum) for 48 h as demonstrated previously (12). Primary cultured human aortic smooth muscle cells (Clonetics, San Diego, CA) were grown in smooth muscle growth medium (Clonetics) containing 5.5 mM glucose, 5% fetal bovine serum, 10 ng/ml human recombinant epidermal growth factor, 390 ng/ml dexamethasone, 50 g/ml gentamicin, and 50 ng/ml amphotericin-B at 37°C in a humidified 5% CO 2 , 95% air atmosphere.…”

Section: Methodsmentioning

confidence: 99%

The Protein Kinase C βII Exon Confers mRNA Instability in the Presence of High Glucose Concentrations

Patel

Eichler

Chappell

et al. 2003

Journal of Biological Chemistry

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Previous studies showed that short term exposure of cells to high glucose destabilized protein kinase C (PKC) ␤II mRNA, whereas PKC␤I mRNA levels remained unaltered. Because PKC␤ mRNAs share common sequences other than the PKC␤II exon encoding a different carboxyl terminus, we examined PKC␤II mRNA for a cis-acting region that could confer glucose-induced destabilization. A ␤-globin/growth hormone reporter con struct containing the PKC␤II exon was transfected into human aorta and rat vascular smooth muscle cells (A10) to follow glucose-induced destabilization. Glucose (25 mM) exposure destabilized PKC␤II chimeric mRNA but not control mRNA. Deletion analysis and electrophoretic mobility shift assays followed by UV cross-linking experiments demonstrated that a region introduced by inclusion of the ␤II exon was required to confer destabilization. Although a cis-acting element mapped to 38 nucleotides within the ␤II exon was necessary to bestow destabilization, it was not sufficient by itself to confer complete mRNA destabilization. Yet, in intact cells antisense oligonucleotides complementary to this region blocked glucose-induced destabilization. These results suggest that this region must function in context with other sequence elements created by exon inclusion involved in affecting mRNA stability. In summary, inclusion of an exon that encodes PKC␤II mRNA introduces a cisacting region that confers destabilization to the mRNA in response to glucose.Protein kinase C (PKC), 1 a serine/threonine kinase, comprises a family of 12 isozymes that have been implicated in signaling pathways affecting cellular processes such as cell proliferation and differentiation, apoptosis, tumor promotion, transcriptional activation, and hormone production (1). The PKC isozymes exhibit differential cellular distribution and substrate specificity. The conventional PKC isozymes, which are Ca 2ϩ -dependent and activated by phospholipid and diacylglycerol, include PKC␣, PKC␤I, PKC␤II, and PKC␥. PKC␤I and PKC␤II are encoded by the same gene, and are translated from alternatively spliced products of PKC␤ pre-mRNA. The inclusion of the PKC␤II exon in the 3Ј-region through alternative splicing results in the PKC␤II mRNA. This pattern of splicing generates a stop codon at the ␤II-␤I boundary such that the ␤I exon, with its coding sequence and 3Ј-UTR, now becomes the 3Ј-UTR of PKC␤II mRNA (Fig. 1). As a result, the PKC␤I and PKC␤II mRNAs differ only by the sequence of the included PKC␤II exon, and the proteins differ only by their carboxylterminal 50 -52 amino acids, respectively (1).We have previously shown that acute hyperglycemia downregulates PKC␤II, but not PKC␤I, at the mRNA and protein levels in vascular smooth muscle cells (2). To understand the mechanism by which elevated glucose down-regulates PKC␤II expression in vascular smooth muscle cells, earlier studies were carried out to determine at what level suppression of PKC␤II expression occurred. These studies clearly demonstrated that high glucose (10 -30 mM), at levels that commonly occ...

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Physiological quiescence in plasma‐derived serum: Influence of platelet‐derived growth factor on cell growth in culture

Cited by 128 publications

References 20 publications

Phenotype-dependent response of cultured aortic smooth muscle to serum mitogens.

Phenotype-dependent response of cultured aortic smooth muscle to serum mitogens.

Active proliferation of Rous sarcoma virus-infected, but not normal, chicken heart mesenchymal cells in culture medium of physiological composition.

The Protein Kinase C βII Exon Confers mRNA Instability in the Presence of High Glucose Concentrations

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