Visualizing tropoelastin in a long-term human elastic fibre cell culture model
| Autor: | Frank Fischer, M. Halm, Horst Wenck, Katja Schenke-Layland, Sören Jaspers |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Materials science Confocal Fibrillin-1 Gene Expression Context (language use) Real-Time Polymerase Chain Reaction Article law.invention Cell Line Transforming Growth Factor beta1 03 medical and health sciences Tissue engineering Bacterial Proteins Confocal microscopy law Tropoelastin Microscopy Humans Multidisciplinary Microscopy Confocal 030102 biochemistry & molecular biology biology Super-resolution microscopy Anatomy Fibroblasts Elastic Tissue Elastin Luminescent Proteins 030104 developmental biology Microscopy Fluorescence biology.protein Biomedical engineering |
| Zdroj: | Scientific Reports |
| ISSN: | 2045-2322 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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