Structural comparisons of fibronectins isolated from early and late passage cells

Sorrentino, Joseph A.; Millis, Albert J.T.

doi:10.1016/0047-6374(84)90155-6

Cited by 28 publications

(5 citation statements)

References 26 publications

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“…Cell-associated fibronectin has been reported to be reduced (Vogel et al, 1981) or coarser and less well organized (Edick and Millis, 1984) on the surface of high passage compared to low passage cells. In addition, the fibronectin produced and secreted into the medium by high passage cells appears to be structurally and functionally different (Chandrasekhar et al, 1983;Sorrentino and Millis, 1984). On an RNA level, a 3-4-fold increase in fibronectin mRNA in high passage human fibroblasts has been observed (Kleinsek, 1989;Seshadri and Campisi, 1989;Kumazaki et al, 1991), observations that agree well with our data on fibronectin mRNA levels in Syrian hamster fibroblasts at the end of their proliferative life span in vitro.…”

Section: Discussionsupporting

confidence: 89%

Differential extracellular matrix gene expression by fibroblasts during their proliferative life span in vitro and at senescence

Choi

Olsen

Cook

et al. 1992

Journal Cellular Physiology

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Increasing evidence supports the idea that the finite proliferative life span of normal fibroblasts is a differentiation-like phenomenon. If this were correct, an ordered sequence of differential gene expression should be associated with the in vitro progression of cells from low passage to high passage (senescence). To define the pattern of expression of fibroblast differentiation-associated genes during this in vitro progression, we have determined the temporal pattern of expression of extracellular matrix (ECM) genes in Syrian hamster dermal fibroblasts as a function of passage level and percentage of proliferative life span in vitro. Steady-state mRNA levels were determined by Northern and dot blot analyses of total cellular RNA hybridized with cDNA probes specific for fibronectin, procollagen alpha 1III, and procollagen alpha 1I. Cells were analyzed at 24 hr postconfluence to minimize the presence of actively proliferating cells, and because maximal levels of fibronectin, alpha 1III, and alpha 1I mRNAs were observed 24 hr postconfluence. Unique, multiphasic patterns of expression of each of these ECM components were observed as the cells progressed from low passage to high passage. As the cells reached midhigh passage, fibronectin mRNA levels increased. This midpassage increase in fibronectin was followed by an increase in the level of alpha 1III mRNA as the cells reached the end of their in vitro proliferative life span, and then alpha 1I when the cells entered the postmitotic senescent phase, at which time the level of fibronectin mRNA also declined. A similar overlapping cascade pattern of up-regulation of these genes is seen during development and wound repair. This suggests that as cultured fibroblasts reach the end of their proliferative life span, they reinitiate a gene expression program used in tissue development and repair.

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Section: Discussionsupporting

confidence: 89%

Differential extracellular matrix gene expression by fibroblasts during their proliferative life span in vitro and at senescence

Choi

Olsen

Cook

et al. 1992

Journal Cellular Physiology

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“…6b,c) that was thought to be fibronectin. Observations in our lab West et al, in preparation) and by others (Chandrasekhar et al, 1983;Edick and Millis, 1984;Sorrentino and Millis, 1984) had shown there were some functional and structural differences between the fibronectin molecules secreted by senescent and young cells in culture. To confirm that the high molecular weight bands observed were fibronectin, two different analyses were performed.…”

Section: Identification Of Antigens As Fibronectinmentioning

confidence: 82%

Novel monoclonal antibodies identify antigenic determinants unique to cellular senescence

Porter

Pereira‐Smith

Smith

1990

Journal Cellular Physiology

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Normal human diploid fibroblasts exhibit a limited lifespan in vitro and are used as a model to study in vivo aging. Monoclonal antibodies were generated against partially purified surface membranes from human diploid fibroblasts at the end of their lifespan (senescent). Three hybridomas were isolated that secreted antibodies reacting to cellular determinants expressed specifically on senescent human fibroblasts of different origin, including neonatal foreskin, embryonic lung, and adult skin punch biopsy, but not expressed on matched young cells. The antibodies did not bind to immortal human cells and normal young cells made reversibly nondividing, indicating the antigens are not expressed in cells that are not senescent. The antibodies identified senescent cells in a mixed cell population and expression of the senescent cell antigens correlated strongly with the cells inability to synthesize DNA at the onset of senescence. The antigens appeared to be cell surface or extracellular matrix associated, and the epitopes were destroyed by mild trypsin treatment. Western analysis indicated all three antibodies reacted with fibronectin. Though the antigenic determinants on the fibronectin molecule were not accessible in the intact young cell, the epitopes were present in fibronectin extracted from both senescent and young cells, as well as purified human plasma fibronectin. These antibodies and the senescent specific expression of the antigens provide powerful tools to investigate the mechanisms leading to in vitro senescence. This may enable us to investigate directly the relationship between cellular aging and aging of the individual.

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“…However, senescent fibroblasts lose their ability to produce collagen and produce high levels of procollagenase and low levels of the tissue inhibitors of metalloproteinases .29 Also, fibronectin from senescent fibroblasts is structurally different from fibronectin isolated from early passage cells and is not able to bind to collagen. 30,31 The potential for senescent ulcer fibroblasts to respond to growth factors within the confines of the margin is an important question that we are cur rently examining. Because most studies suggest that senescent fibroblasts are irreversibly locked into the G1 phase of the cell cycle, 32,33 repair of chronic pressure ulcers may involve fibroblasts that were not fully senescent.…”

Section: Vande Berg Et Al 47mentioning

confidence: 99%

Fibroblast senescence in pressure ulcers

Berg

Rudolph

Hollan

et al. 1998

Wound Repair Regeneration

117

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Pressure ulcers appear as localized chronic wounds in the middle of normal functioning skin. This study focuses on pressure ulcer fibroblasts cultured from the ulcer bed, ulcer margin, and normal, nonulcerated skin adjacent to the wound. From these areas we show that pressure ulcer fibroblasts become prematurely senescent. Verification of senescence was based on failure of cell populations to undergo a 0.5 population doubling after 1 week in culture, light microscopic appearance of late-passage senescent cells in culture, light microscopic verification of senescence in vivo and in vitro by anti-terminin staining, and ultrastructural identification of senescent fibroblasts by anti-terminin colloidal gold labeling. Although not all ulcer fibroblasts are senescent, there is a range of proliferative ability. The senescent phenotype of ulcer fibroblasts remains intact in vitro as these cells remain viable but are unable to complete DNA synthesis. Fibroblast senescence is not uniform in chronic wounds but varies within areas of the ulcer bed and from patient to patient. Although ulcer fibroblasts exhibit limited proliferative ability, fibroblasts from the ulcer margin and adjacent normal skin show a continued ability to divide. However, in all areas of the wound, the mean generation time of the fibroblast population is longer than commonly observed for cultured human diploid fibroblasts. At first passage, the mean generation time for fibroblasts from all patients is significantly different (p = .001, analysis of variance) among fibroblasts from the ulcer bed (4.2 2.2 days), ulcer margin (2.9 +/- 1.3 days), and adjacent normal skin (1.9 +/- 0.6 days). Analysis of the three cell groups by the post hoc Tukey test, using corrected p values shows that the difference between mean generation times among fibroblast populations from adjacent normal skin and ulcer bed are significantly different (p < 0.05), whereas the difference between fibroblasts from adjacent normal skin and ulcer margin and ulcer margin and ulcer bed are not significantly different (p > 0.05). These data may help to explain the poor response of certain pressure sores to aggressive medical treatment.

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Structural comparisons of fibronectins isolated from early and late passage cells

Cited by 28 publications

References 26 publications

Differential extracellular matrix gene expression by fibroblasts during their proliferative life span in vitro and at senescence

Differential extracellular matrix gene expression by fibroblasts during their proliferative life span in vitro and at senescence

Novel monoclonal antibodies identify antigenic determinants unique to cellular senescence

Fibroblast senescence in pressure ulcers

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