Culture of human dermal fibroblasts in collagen gels: Modulation of interleukin 1-induced prostaglandin E2 synthesis by an extracellular matrix

In the dermis, fibroblasts play an important role in the turnover of the dermal extracellular matrix. Collagen I and III, the most important dermal proteins of the extracellular matrix, are progressively altered during ageing and diabetes. For mimicking diabetic conditions, the cultured human dermal fibroblasts were incubated with increasing amounts of AGE-modified BSA andD-glucose for 24 hours. The expression of procollagenα2(I) and procollagenα1(III) mRNA was analyzed by quantitative real-time PCR. Our data revealed that the treatment of fibroblasts with AGE-modified BSA upregulated the expression of procollagenα2(I) and procollagenα1(III) mRNA in a dose-dependent manner. High glucose levels mildly induced a profibrogenic pattern, increasing the procollagenα2(I) mRNA expression whereas there was a downregulation tendency of procollagenα1(III) mRNA.

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Detection of Interleukin 1 with Human Dermal Fibroblasts

Immunobiology ◽

10.1016/s0171-2985(89)80023-3 ◽

1989 ◽

Vol 179 (2-3) ◽

pp. 283-291 ◽

Cited By ~ 26

Author(s):

Harald Loppnow ◽

Hans-Dieter Flad ◽

Iris Dürrbaum ◽

Joachim Musehold ◽

Rudolf Fetting ◽

...

Keyword(s):

Interleukin 1 ◽

Dermal Fibroblasts ◽

Human Dermal Fibroblasts

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Comparative Effects of Interleukin-1 and Tumor Necrosis Factor-α on Collagen Production and Corresponding Procollagen mRNA Levels in Human Dermal Fibroblasts

Journal of Investigative Dermatology ◽

10.1111/1523-1747.ep12462185 ◽

1991 ◽

Vol 96 (2) ◽

pp. 243-249 ◽

Cited By ~ 83

Author(s):

Alain Mauviel ◽

Jyrki Heino ◽

Veli-Matti Kähäri ◽

Daniel-Jean Hartmann ◽

Gérard Loyau ◽

...

Keyword(s):

Tumor Necrosis Factor ◽

Tumor Necrosis ◽

Tumor Necrosis Factor Α ◽

Interleukin 1 ◽

Dermal Fibroblasts ◽

Mrna Levels ◽

Human Dermal Fibroblasts ◽

Collagen Production ◽

Factor Α ◽

Necrosis Factor

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Interleukin-1 Beta Promotes the Expression of Prox-1 and Vascular Endothelial Growth Factor Receptor Type-3 in Adult Human Dermal Fibroblasts

Blood ◽

10.1182/blood.v120.21.4738.4738 ◽

2012 ◽

Vol 120 (21) ◽

pp. 4738-4738

Author(s):

Matsuo Takuji ◽

Ryosuke Shirasaki ◽

Haruko Tashiro ◽

Yoko Oka ◽

Tadashi Yamamoto ◽

...

Keyword(s):

Morphological Changes ◽

Interleukin 1 ◽

Growth Factor Receptor ◽

Dermal Fibroblasts ◽

Vascular Endothelial ◽

Human Dermal Fibroblasts ◽

Interleukin 1 Beta ◽

Lymphatic Duct ◽

Type 3 ◽

Endothelial Growth Factor

Abstract Abstract 4738 Background and Aims: We recently reported that human interleukin 1 beta (IL-1 beta) stimulates bone-marrow stromal myofibroblasts to express a hematopoietic stem cell marker, CD34 (16th EHA, and 53rdASH meeting). To characterize precisely the effects of IL-1 beta to the stromal cells, adult human dermal fibroblasts (HDF) were cultured with IL-1 beta, and the morphological changes and molecular expressions were observed. Materials and Methods: HDF were purchased, and cultured in knockout DMEM medium with 15% KSR with or without recombinant human IL-1 beta for 3 weeks. The morphological changes and the expression of specific hematopoiesis-related genes were analyzed time-dependently. The concentration of hematopoietic growth factors in the culturing supernatants was also measured. Results: When HDF were cultured with human IL-1 beta for 3 weeks, the cellular morphology changed to the filamentous appearance. RT-PCR analyses revealed that cells expressed Prox-1, a master molecule for lymphatic duct neogenesis, and also vascular endothelial growth factor receptor (VEGFR) type-3. Smad7 and PAI1 were also expressed; however, LYVE1, a marker for lymphatic vascular endothelial cell, was not induced. VEGF-C production was also demonstrated at RNA and protein levels, and VEGF-C and VEGFR type-3 system played an important role in morphological changes and cell-proliferation. Discussion: We previously reported that when HDF were cultured in DMEM/F12 medium with IL-1 beta and EPO, hematopoietic markers were induced. When HDF were cultured in knockout DMEM, a condition for immature cell-growth at ES cell levels, and IL-1 beta, lymphatic duct neogenesis-related genes were activated, and HDF converted their morphology without an introduction of any genes externally. We are now making clear the precise action of IL-1 beta to HDF using microarray analysis. Disclosures: No relevant conflicts of interest to declare.

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Expression of pro-inflammatory markers by human dermal fibroblasts in a three-dimensional culture model is mediated by an autocrine interleukin-1 loop

Biochemical Journal ◽

10.1042/bj20031371 ◽

2004 ◽

Vol 379 (2) ◽

pp. 351-358 ◽

Cited By ~ 29

Author(s):

Daniela KESSLER-BECKER ◽

Thomas KRIEG ◽

Beate ECKES

Keyword(s):

Keratinocyte Growth Factor ◽

Interleukin 1 ◽

Three Dimensional ◽

Dermal Fibroblasts ◽

Human Dermal Fibroblasts ◽

Inflammatory Processes ◽

Cox 2 ◽

Extracellular Matrix Interactions

In vivo, fibroblasts reside in connective tissues, with which they communicate in a reciprocal way. Such cell–extracellular matrix interactions can be studied in vitro by seeding fibroblasts in collagen lattices. Depending upon the mechanical properties of the system, fibroblasts are activated to assume defined phenotypes. In the present study, we examined a transcriptional profile of primary human dermal fibroblasts cultured in a relaxed collagen environment and found relative induction (>2-fold) of 393 out of approx. 7100 transcripts when compared with the same system under mechanical tension. Despite down-regulated proliferation and matrix synthesis, cells did not become generally quiescent, since they induced transcription of numerous other genes including matrix metalloproteinases (MMPs) and growth factors/cytokines. Of particular interest was the induction of gene transcripts encoding pro-inflammatory mediators, e.g. cyclo-oxygenase-2 (COX-2), and interleukins (ILs)-1 and -6. These are apparently regulated in a hierarchical fashion, since the addition of IL-1 receptor antagonist prevented induction of COX-2, IL-1 and IL-6, but not that of MMP-1 or keratinocyte growth factor (KGF). Our results suggest strongly that skin fibroblasts are versatile cells, which adapt to their extracellular environment by displaying specific phenotypes. One such phenotype, induced by a mechanically relaxed collagen environment, is the ‘pro-inflammatory’ fibroblast. We propose that fibroblasts that are embedded in a matrix environment can actively participate in the regulation of inflammatory processes.

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