Tissue-specific differences in cultured human diploid fibroblasts

Schneider, Erasmus; MITSUI, Y; Au, Kam; Shorr, Stuart S.

doi:10.1016/0014-4827(77)90320-2

Cited by 11 publications

(13 citation statements)

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“…Compared to lung fibroblasts (Holz et al 2004), values were lower in skin fibroblasts, in line with previous findings (Schneider et al 1977). Moreover, the proportion of skin fibroblasts positive for β-gal was not significantly elevated in emphysema.…”

Section: Accepted Manuscriptsupporting

confidence: 89%

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In contrast to lung fibroblasts – no signs of senescence in skin fibroblasts of patients with emphysema

Müller

Paasch

Feindt

et al. 2008

Experimental Gerontology

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Section: Accepted Manuscriptsupporting

confidence: 89%

“…Foetal skin fibroblasts appear to show a lower proliferation rate than lung fibroblasts (Schneider et al 1977). In line with this, the ratio of doubling times between lung and skin was <1 in control patients, whereas in emphysema the slow proliferation of lung fibroblasts raised this ratio to values >1.…”

Section: Accepted Manuscriptsupporting

confidence: 66%

In contrast to lung fibroblasts – no signs of senescence in skin fibroblasts of patients with emphysema

Müller

Paasch

Feindt

et al. 2008

Experimental Gerontology

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“…2 compares the O'Farrell gel patterns of total cellular polypeptides from a skin fibroblast line (GM738) and a lung fibroblast line (MRC5) by double-label autoradiography. Skin and lung fibroblasts may not be metabolically identical (16), and their O'Farrell gel patterns reveal many more quantitative differences than were seen when any two lung fibroblast lines were compared (Fig. 1B).…”

Section: Cellmentioning

confidence: 96%

Human heterozygosity: a new estimate.

McConkey¹,

Taylor²,

Phan³

1979

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

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Several hundred polypeptides from four human diploid fibroblast cell lines were compared by high-resolution two-dimensional polyacrylamide gel electrophoresis and double-label autoradiography under conditions where allelic products that differ by a single charged amino acid would be distinguished. The average heterozygosity represented by this set of gene products appears to be less than 1% for changes involving charged amino acids.The nature and extent of genetic diversity among humans is a matter of great theoretical and practical significance. Our current views on this subject depend heavily on the summaries by Harris and colleagues (1, 2). Of the 87 (6)(7)(8). These facts suggested to one of us several years ago that an attempt to measure heterozygosity by comparison of O'Farrell gel patterns of polypeptides from cells of different individuals would be fruitful. Side-by-side comparison of complex patterns, however, is fatiguing and inaccurate. The problem of finding a few spots unique to one two-dimensional gel pattern among hundreds of spots common to both patterns in any pairwise comparison has been solved by the development of double-label autoradiography (9). This method permits the simultaneous electrophoresis of 3H-labeled and '4C-labeled protein mixtures on the same gel, followed by a two-step autoradiographic procedure that identifies the polypeptides that are unique to the 3H-labeled preparation. We have used this technique to compare approximately 400 fibroblast polypeptides from several individuals. We find that the heterozygosity of the genes represented by this set of polypeptides is significantly less than one would have expected from the enzyme surveys (1, 2). (2.5 ,Ci/ml, 300 Ci/mol) (1 Ci = 3.7 X 1010 becquerels). Cells were harvested by scraping with a rubber policeman, rinsed with 0.14 M NaCl/0.01 M sodium phosphate, pH 7.0, and pelleted in a conical tube which was then carefully wiped dry. Fresh or frozen cell pellets were lysed in freshly made 10 M urea, with vigorous mixing on a Vortex. The protein concentration of a 5-to 10-,ul sample of homogenate was measured with the Bio-Rad Reagent (bulletin 1069), and 1% sodium dodecyl sulfate/10 M urea was added so that the final mass ratio of detergent to protein was 1.5:1. If necessary, 10 M urea was added so that the final protein concentration did not exceed 1 mg/ml. The homogenate was then made 0.1 M in acetic acid (final pH 4.0-4.2) and 5 mM in ZnSO4; SI nuclease (Sigma or P-L Biochemicals) was added at 2 units of enzyme per 1ug of homogenate protein, and the mixture was incubated at 450C for 5 min, then chilled. An equal volume of 1 M acetic acid/1% 2-mercaptoethanol was added, followed by 10 homogenate volumes of cold acetone, and the proteins were allowed to precipitate at -20°C overnight. The precipitate was collected by centrifugation at 10,000 X g for 15 min. MATERIALS AND METHODS CellSamples were dissolved in O'Farrell's buffer (5) and centrifuged for 10 min at 10,000 X g at 25°C just before electrofocusing gels were lo...

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“…Fibroblast heterogeneity has been very well documented over the last two decades with respect to a variety of parameters including differences in proliferation rates, 55 responses to prostaglandins, 56 collagen synthesis, 57 cytokine production 58 and cytokine receptors. 59 Phenotypic diversity also exists within fibroblast populations, along with differences in morphology 60 and cell size.…”

Section: Deep Dermal Fibroblasts Dictate Hts J Wang Et Almentioning

confidence: 99%

Deep dermal fibroblasts contribute to hypertrophic scarring

Wang

Dodd

Shankowsky

et al. 2008

Laboratory Investigation

185

140

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Hypertrophic scar (HTS) following thermal injury is a dermal fibroproliferative disorder that leads to considerable morbidity. The development of HTS involves numerous cell types and cytokines with dermal fibroblasts being a key cell. We have previously reported that the phenotype of fibroblasts isolated from HTS was altered compared to fibroblasts from normal skin. In this study, normal skin was horizontally sectioned into five layers using a dermatome from which fibroblasts were isolated and cultured. Cells from the deeper layers were observed to proliferate at a slow rate, but were morphologically larger. In ELISA and FACS assays, cells from the deeper layers produced more TGF-b1 and TGF-b1 producing cells were higher. In quantitative RT-PCR, the cells from the deeper layers had higher CTGF and HSP47 mRNA levels compared to those from superficial layers. In western blot, FACS and collagen gel assays, fibroblasts from the deeper layers produced more a-smooth muscle actin (a-SMA), had higher a-SMA positive cells and contracted collagen gels more. Fibroblasts from the deeper layers were also found to produce more collagen, but less collagenase by mass spectrometry and collagenase assay. Interestingly, cells from the deeper layers also produced more of the proteoglycan, versican, but less decorin. Taken together, these data strongly demonstrate that fibroblasts from the deeper layers of the dermis resemble HTS fibroblasts, suggesting that the deeper layer fibroblasts may be critical in the formation of HTS. Hypertrophic scarring is a common fibroproliferative disorder of the human dermis characterized by erythematous, raised, pruritic lesions of the healing skin, which is usually following thermal and other injuries that involve the deep dermis. 1 These lesions lead to scarring that compromises the appearance of healing skin and are commonly associated with contractures that limit movement and function of involved joints and facial features. As such, HTS is the principal factor that contributes to the prolonged and often uncomfortable rehabilitation period for thermally injured patients, particularly those who have survived large lifethreatening injuries, children, and individuals of dark-skinned races, in whom such scarring occurs more commonly. 2 The undesirable physical properties of HTS tissue can be attributed to the presence of a large amount of extracellular matrix that is of altered composition and organization, compared to normal dermis or mature scar. This matrix is the product of a dense population of fibroblasts, maintained in a hyperactive state by inflammatory cytokines such as TGF-b and other factors, some of which may be physical in origin. 3 Although the molecular and cellular events that lead to HTS have been extensively studied, the pathogenesis of this condition is still not well understood, making treatment difficult.Key cells involved in HTS are the dermal fibroblasts. Dermal fibroblasts are a dynamic and diverse population of cells whose function in skin and HTS in many respects remain unkn...

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Tissue-specific differences in cultured human diploid fibroblasts

Cited by 11 publications

References 0 publications

In contrast to lung fibroblasts – no signs of senescence in skin fibroblasts of patients with emphysema

In contrast to lung fibroblasts – no signs of senescence in skin fibroblasts of patients with emphysema

Human heterozygosity: a new estimate.

Deep dermal fibroblasts contribute to hypertrophic scarring

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