title: Visualizing tropoelastin in a long-term human elastic fibre cell culture model.
abstract: Elastin is an essential protein found in a variety of tissues where resilience and flexibility are needed, such as the skin and the heart. When aiming to engineer suitable implants, elastic fibres are needed to allow adequate tissue renewal. However, the visualization of human elastogenesis remains in the dark. To date, the visualization of human tropoelastin (TE) production in a human cell context and its fibre assembly under live cell conditions has not been achieved. Here, we present a long-term cell culture model of human dermal fibroblasts expressing fluorescence-labelled human TE. We employed a lentiviral system to stably overexpress Citrine-labelled TE to build a fluorescent fibre network. Using immunofluorescence, we confirmed the functionality of the Citrine-tagged TE. Furthermore, we visualized the fibre assembly over the course of several days using confocal microscopy. Applying super resolution microscopy, we were able to investigate the inner structure of the elastin-fibrillin-1 fibre network. Future investigations will allow the tracking of TE produced under various conditions. In tissue engineering applications the fluorescent fibre network can be visualized under various conditions or it serves as a tool for investigating fibre degradation processes in disease-in-a-dish-models.
authors: Research and Development, Beiersdorf AG, Unnastr. 48, Hamburg, Germany.
authors: M
authors: M
authors: Halm
authors: Dept. of Women's Health, Research Institute for Women's Health, Eberhard-Karls-University Tübingen, Silcherstr. 7/1, 72076 Tübingen, Germany.|Dept. of Cell and Tissue Engineering, Fraunhofer Institute for Interfacial Engineering and Biotechnology (IGB), Nobelstr. 12, 70569 Stuttgart, Germany.|Dept. of Medicine/Cardiology, Cardiovascular Research Laboratories, David Geffen School of Medicine at UCLA, 675 Charles E. Young Drive South, MRL 3645, Los Angeles, CA, USA.
authors: K
authors: K
authors: Schenke-Layland
authors: Research and Development, Beiersdorf AG, Unnastr. 48, Hamburg, Germany.
authors: S
authors: S
authors: Jaspers
authors: Research and Development, Beiersdorf AG, Unnastr. 48, Hamburg, Germany.
authors: H
authors: H
authors: Wenck
authors: Research and Development, Beiersdorf AG, Unnastr. 48, Hamburg, Germany.
authors: F
authors: F
authors: Fischer
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D004547:Elastic Tissue / Q000378:metabolism* / Q000648:ultrastructure; D004549:Elastin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000071838:Fibrillin-1 / Q000737:chemistry / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D060888:Real-Time Polymerase Chain Reaction; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology; D014333:Tropoelastin / Q000737:chemistry / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D000071838:Fibrillin-1; D008164:Luminescent Proteins; D053773:Transforming Growth Factor beta1; D014333:Tropoelastin; C434479:citrine protein, bacteria; D004549:Elastin