Influence of Increased Extracellular Calcium Concentration and Donor Age on Density-Dependent Growth Inhibition of Human Fibroblasts

Praeger, Frank C.; Gilchrest, Barbara A.

doi:10.3181/00379727-182-42345

Cited by 16 publications

(4 citation statements)

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“…NFFs have the highest growth capacity among fibroblasts cultured from all age groups. 14,15 NFFs from the same donor were used in each experiment to exclude the possibility of donor variability. Neonatal foreskin was obtained within 2 to 6 hours of elective circumcision, and primary cultures of dermal fibroblasts were prepared with techniques described in detail by Stanley et al 16 In brief, biopsy specimens were treated with a 5-minute povidoneiodine bath followed by a 5-minute bath in 70% ethanol.…”

Section: Fibroblast Isolation and Culturementioning

confidence: 99%

The proliferative capacity of neonatal skin fibroblasts is reduced after exposure to venous ulcer wound fluid: A potential mechanism for senescence in venous ulcers

Mendez

Raffetto

Phillips

et al. 1999

Journal of Vascular Surgery

122

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These data suggest that the venous ulcer microenvironment adversely affects young, rapidly proliferating fibroblasts such as NFFs and induces fibroblast senescence. Pro-inflammatory cytokines such as TNF-alpha and TGF-beta1 might be involved in this process. The role of other unknown inhibitory mediators, as well as pro-inflammatory cytokines, in venous ulcer development and impaired healing must be considered.

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Section: Fibroblast Isolation and Culturementioning

confidence: 99%

The proliferative capacity of neonatal skin fibroblasts is reduced after exposure to venous ulcer wound fluid: A potential mechanism for senescence in venous ulcers

Mendez

Raffetto

Phillips

et al. 1999

Journal of Vascular Surgery

122

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“…Cell source Normal human fibroblast cultures were established from neonatal foreskin and adult specimens as previously described (Gilchrest, 1980;Praeger and Gilchrest, 1986). Cells at passage levels 2 4 were processed in one of two ways: 1) Cells were inoculated at low density (lo3 celldcm') or high density (lo4 cells/ cm2) in DME (Gibco, Grand Island, NY) supplemented with 10% FBS (Gibco).…”

Section: Methodsmentioning

confidence: 99%

Dissociation of proto‐oncogene induction from growth response in normal human fibroblasts

Yaar

Peacocke

Cohen

et al. 1990

Journal Cellular Physiology

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Proto-oncogenes are cellular homologues of viral oncogenes that are known to be associated with regulation of growth and differentiation. The c-myc and c-fos proto-oncogenes have been extensively studied by using established cell lines but to a lesser extent by using normal cells. Using physiologic growth modulators, we have shown that mitotic stimulation of normal human dermal fibroblasts is associated with induction of c-myc and c-fos, but that growth inhibition of these cells is not necessarily accompanied by their down-regulation. When treated with both serum and interferon-alpha during quiescence, fibroblasts were delayed in their progress into the S-phase of the cell cycle as compared to cells treated with serum alone and displayed substantial growth inhibition as measured by cell number at the end of 1 week. However, this growth inhibition was not preceded by down-regulation or delay in induction of c-myc and c-fos mRNA. The above studies suggest that in normal fibroblasts growth inhibition is not necessarily dependent on down-regulation of transcription of either c-myc or c-fos and that interferon may act to inhibit cell growth either through a post-transcriptional effect on cellular proto-oncogenes necessary for cell proliferation or through induction of other, as yet unrecognized gene(s) associated with growth arrest.

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“…In fact, serum-stimulated confluent foreskin-derived fibroblasts in the presence of additional calcium chloride (CaC12) show an increase in cell yield that requires multiple rounds of cell division by a sizable fraction of the cell culture (5). Previous studies by other investigators using a wide variety of experimental approaches have demonstrated similar effects for calcium precipitate formed by the introduction of pyrophosphate (PPJ (7) or the insoluble calcium crystal hydroxyapatite (HA) (8) or lanthanum chloride (LaC13), even when presented to cells only briefly under cell culture conditions adjusted so as to prevent precipitation (9).…”

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confidence: 99%

Calcium, Lanthanum, Pyrophosphate, and Hydroxyapatite: A Comparative Study in Fibroblast Mitogenicity

Praeger

Gilchrest

1989

Experimental Biology and Medicine

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Calcium-containing crystals and elevated levels of calcium chloride (CaCI,) and lanthanum chloride (LaCI,) have been previously reported to enhance the proliferative activity of cultured fibroblasts. We have investigated the relative mitogenicity of these agents, whether they function via precipitation on the cell surface and whether they interact with one another. Confluent cultures of newborn foreskin fibroblasts provided with fresh medium containing 10% fetal bovine serum (FBS) in the presence of hydroxyapatite (HA), pyrophosphate (PP,), LaCI, (La), or additional CaCI, (Ca) were all stimulated more than control cultures provided with fresh medium and 10% FBS alone as assessed by cell counts 5 days later. Increases in cell yield above the original confluent cell density were 316% for La, 271% for Ca, 189% for HA, 131% for PP,, and 45% for controls. Addition of fresh medium containing 10% FBS and epidermal growth factor or fresh medium containing 20% FBS as additional points of reference yielded increases of 204 and 107%, respectively, over original confluent density. Stimulation induced by La or Ca was significantly greater (P c 0.001) than the stimulations induced by each of the other treatments. The same treatments added to confluent cultures without a change of medium also renewed mitotic activity, with La and Ca again the most mitogenic and approximately doubling the pretreatment cell yields. Cultures incubated in an inverted position to avoid cell contact with precipitates in the medium were also stimulated by La and Ca, but not by HA and PP,. When added to confluent cultures simultaneously supplemented with optimal additional Ca, La decreased Day 5 cell yields in a dose-dependent manner at low concentrations (0.03-0.2 mM) but increased cell yields over those obtained with 0.2 mM LaCI, again in a dose-dependent manner at higher concentrations. Thus, while HA and PP, act via precipitation on the cell surface, the more mitogenic agents La and Ca function in solution and appear to stimulate cell division by different nonadditive mechanisms. These findings suggest multiple mechanisms of membrane participation in mitogen responsiveness and in density-dependent inhibition of growth. [P.S.E.B.M. 1989[P.S.E.B.M. , Vol 1901 ensity-dependent inhibition of growth is a characteristic behavior of cultured diploid fibro-D blasts that is altered by cellular aging in vitro ( I, 2) but not expressed by cultured malignant or malig- nantly transformed cells (3, 4). Hence, although its mechanism is poorly understood, density-dependent growth inhibition can be assumed to reflect important aspects of cellular growth regulation.Recent studies with normal human fibroblasts show that elevating extracellular calcium concentration 'This work was supported by the Agricultural Research Service

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Influence of Increased Extracellular Calcium Concentration and Donor Age on Density-Dependent Growth Inhibition of Human Fibroblasts

Cited by 16 publications

References 0 publications

The proliferative capacity of neonatal skin fibroblasts is reduced after exposure to venous ulcer wound fluid: A potential mechanism for senescence in venous ulcers

The proliferative capacity of neonatal skin fibroblasts is reduced after exposure to venous ulcer wound fluid: A potential mechanism for senescence in venous ulcers

Dissociation of proto‐oncogene induction from growth response in normal human fibroblasts

Calcium, Lanthanum, Pyrophosphate, and Hydroxyapatite: A Comparative Study in Fibroblast Mitogenicity

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