Control of growth and expression of differentiated functions of mature hepatocytes in primary culture.

Nakamura, Toshikazu; Ichihara, Akira

doi:10.1247/csf.10.1

Cited by 58 publications

(19 citation statements)

References 34 publications

(6 reference statements)

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“…A striking characteristic of the response to EGF was its heterogeneity from cell to cell, which is evident for each parameter studied (lag time, number and duration of the single transients, duration of the oscillatory response, interspike interval). A large variability has already been described for the Ca 2þ oscillations induced by EGF in rat hepatocytes (Tanaka et al, 1992) and human A431 cells (Cheyette and Gross, 1991) and could be accounted for by a number of mechanisms: (1) heterogeneity in the phase of the cell cycle, which could shift from G 0 towards G 1 under the dispersed conditions of the primary culture (Nakamura and Ichihara, 1985) and cause different rates of sphingosine kinase activity between nonoscillating (in G 0 ) and oscillating (in G 1 ) cells, as proposed for the Ca 2þ signals induced by PDGF-BB in transformed oligodentrocytes Miller, 1996, 1999); (2) wide variation from cell to cell, even from the same coverslip, in the number of EGF-R (Cheyette and Gross, 1991) or in their binding properties (Chung et al, 1997); (3) variability in the expression of low-affinity EGF-R, which might be responsible for EGF-elicited Ca 2þ influx (Defize et al, 1989) and, therefore, for the maintained response to the growth factor (see below). The first mechanism seems to be ruled out by preliminary experiments in which the application of 10, 20, and 25 ng/ml EGF to serum-deprived CMEC resulted in patterns of response similar to those described in presence of 10% serum in the growing medium (see Materials and Methods).…”

Section: Discussionmentioning

confidence: 99%

Epidermal growth factor induces intracellular Ca²⁺ oscillations in microvascular endothelial cells

Moccia

Berra‐Romani

Tritto

et al. 2002

Journal Cellular Physiology

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An increase in intracellular Ca(2+) concentration ([Ca(2+)](i)) may play a role in the proliferative effect of several growth factors. In this study, the changes in [Ca(2+)](i) elicited by epidermal growth factor (EGF) in rat cardiac microvascular endothelial cells (CMEC) have been investigated by using fura-2 conventional and confocal microscopy. A large heterogeneity in the latency and in the pattern of the Ca(2+) response was found at each dose of EGF (2.5-100 ng/ml), whereas some cells displayed a non-oscillatory behavior and others exhibited a variable number of Ca(2+) oscillations. On average, the fraction of responsive cells, the total number of oscillations and the duration of the Ca(2+) signal were higher at around 10 ng/ml EGF, while there was no dose-dependence in the lag time and in the amplitude of the [Ca(2+)](i) increase. EGF-induced Ca(2+) spikes were abolished by the tyrosine kinase inhibitor genistein, but not by its inactive analogue daidzein, and by the phospholipase C blocker NCDC. Only 1-2 transients could be elicited in Ca(2+)-free solution, while re-addition of extracellular Ca(2+) recovered the spiking activity. The oscillatory signal was prevented by the SERCA inhibitor thapsigargin and abolished by the calcium entry blockers Ni(2+) and La(3+). Moreover, EGF-induced Ca(2+) transients were abolished by the InsP(3) receptor blocker caffeine, while ryanodine was without effect. Confocal imaging microscopy showed that the Ca(2+) response to EGF was localized both in the cytoplasm and in the nucleus. We suggest that EGF-induced [Ca(2+)](i) increase may play a role in the proliferative action of EGF on endothelial cells.

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Section: Discussionmentioning

confidence: 99%

Epidermal growth factor induces intracellular Ca²⁺ oscillations in microvascular endothelial cells

Moccia

Berra‐Romani

Tritto

et al. 2002

Journal Cellular Physiology

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“…One unit of activity is defined as the quantity of EGF required for half-maximal stimulation of DNA synthesis of adult rat hepatocytes in primary culture. 3. Purification by heparin-Sepharose CL-6B chromatography of partially purified HGF.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, several groups including ours found that adult rat hepatocytes in primary culture, which retain many of their in vivo liver functions, can proliferate when cultured at low density in medium containing insulin and epidermal growth factor (EGF) (1)(2)(3)(4)(5)(6). Thus, primary cultures of adult rat hepatocytes are a suitable in vitro system for use in studies on the humoral factor for hepatocyte growth.…”

mentioning

confidence: 99%

Purification and characterization of a growth factor from rat platelets for mature parenchymal hepatocytes in primary cultures.

Nakamura¹,

Teramoto²,

Ichihara³

1986

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

544

315

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A growth factor (HGF) stimulating DNA synthesis of adult rat hepatocytes in primary culture was found in rat platelets. HGF was purified from rat platelets to homogeneity by a three-step procedure: stimulation of its release from platelets by thrombin, cation-exchanger fast protein liquid chromatography on a Mono S column, and heparin-Sepharose chromatography. HGF was clearly distinguishable from the platelet-derived growth factor (PDGF) by fast protein liquid chromatography. HGF was a heat-and acid-labile cationic protein that was inactivated by reduction with dithiothreitol. Its molecular mass was estimated to be 27 kDa by NaDodSO4/PAGE and its amino acid composition was very different from that of PDGF. The purified HGF stimulated DNA synthesis in adult rat hepatocytes at 2 ng/ml and was maximally effective at 20 ng/ml; its effect was additive or synergistic with those of insulin and EGF, depending on their combinations. HGF did not stimulate DNA synthesis of Swiss 3T3 cells, while PDGF did not stimulate that of hepatocytes. Thus, HGF showed clearly different cell specificity from PDGF in its growth-promoting activities. These findings indicate that HGF is a growth factor in platelets for mature hepatocytes.

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“…Specific growth factors for liver have not yet been unequivocally identified (see Nakamura and Ichihara, 1985), but studies with primary hepatocyte cultures have shown marked hepatotrophic activity associated with platelets (Nakamura and Ichihara, 1985), which cannot be ascribed to PDGF (McGowan, 1986). During liver regeneration after partial hepatectomy, the concentration of prostaglandins E2 and F2, in the serum are increased.…”

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confidence: 99%

Regulation of the proliferation of primary rat hepatocytes by eicosanoids

Skouteris

Ord

Stocken

1988

Journal Cellular Physiology

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DNA synthesis in primary adult rat hepatocyte cultures was promoted by epidermal growth factor (EGF), arachidonic acid, and prostaglandins E2 and F2 alpha (PGE2 and PGF2 alpha). Growth promotion by EGF was blocked by 0.1 mM indomethacin and 1 mM aspirin, without affecting cell viability. If verapamil was present in the medium when EGF was added, the growth response was inhibited. Hepatocytes stimulated by EGF or arachidonic acid released PGE2 and PGF2 alpha into the culture medium. This was diminished if 0.1 mM indomethacin was also in the medium. The importance of autocrine regulation of hepatocyte growth by prostaglandins is discussed.

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Control of growth and expression of differentiated functions of mature hepatocytes in primary culture.

Cited by 58 publications

References 34 publications

Epidermal growth factor induces intracellular Ca²⁺ oscillations in microvascular endothelial cells

Epidermal growth factor induces intracellular Ca²⁺ oscillations in microvascular endothelial cells

Purification and characterization of a growth factor from rat platelets for mature parenchymal hepatocytes in primary cultures.

Regulation of the proliferation of primary rat hepatocytes by eicosanoids

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Control of growth and expression of differentiated functions of mature hepatocytes in primary culture.

Cited by 58 publications

References 34 publications

Epidermal growth factor induces intracellular Ca2+ oscillations in microvascular endothelial cells

Epidermal growth factor induces intracellular Ca2+ oscillations in microvascular endothelial cells

Purification and characterization of a growth factor from rat platelets for mature parenchymal hepatocytes in primary cultures.

Regulation of the proliferation of primary rat hepatocytes by eicosanoids

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Product

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Epidermal growth factor induces intracellular Ca²⁺ oscillations in microvascular endothelial cells

Epidermal growth factor induces intracellular Ca²⁺ oscillations in microvascular endothelial cells