Human fetal and adult epicardial-derived cells: a novel model to study their activation
| Autor: | Asja T. Moerkamp, Thomas J van Brakel, Calinda K. E. Dingenouts, Marie-José Goumans, Esther Dronkers, Kirsten Lodder, Tessa van Herwaarden, Fredrik C. Tengström, Anke M. Smits |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Cell type Pathology medicine.medical_specialty Epithelial-Mesenchymal Transition Organogenesis Human EPDCs Medicine (miscellaneous) Biology Biochemistry Genetics and Molecular Biology (miscellaneous) TGFβ 03 medical and health sciences Fetus Cardiac development In vitro model TGF beta Cell Movement medicine Humans Epithelial–mesenchymal transition Receptor Cells Cultured Tube formation Matrigel Heart development Research Heart Cell Biology Epicardium Cell biology Drug Combinations 030104 developmental biology Epithelial-to-mesenchymal transition embryonic structures Molecular Medicine Proteoglycans Collagen Laminin Stem cell Pericardium Receptors Transforming Growth Factor beta |
| Zdroj: | Stem Cell Research & Therapy Stem Cell Research and Therapy, 7 |
| ISSN: | 1757-6512 |
| DOI: | 10.1186/s13287-016-0434-9 |
| Popis: | Background The epicardium, a cell layer covering the heart, plays an important role during cardiogenesis providing cardiovascular cell types and instructive signals, but becomes quiescent during adulthood. Upon cardiac injury the epicardium is activated, which includes induction of a developmental gene program, epithelial-to-mesenchymal transition (EMT) and migration. However, the response of the adult epicardium is suboptimal compared to the active contribution of the fetal epicardium to heart development. To understand the therapeutic value of epicardial-derived cells (EPDCs), a direct comparison of fetal and adult sources is paramount. Such analysis has been hampered by the lack of appropriate culture systems. Methods Human fetal and adult EPDCs were isolated from cardiac specimens obtained after informed consent. EPDCs were cultured in the presence of an inhibitor of the TGFβ receptor ALK5. EMT was induced by stimulation with 1 ng/ml TGFβ. PCR, immunofluorescent staining, scratch assay, tube formation assay and RT2-PCR for human EMT genes were performed to functionally characterize and compare fetal and adult EPDCs. Results In this study, a novel protocol is presented that allows efficient isolation of human EPDCs from fetal and adult heart tissue. In vitro, EPDCs maintain epithelial characteristics and undergo EMT upon TGFβ stimulation. Although similar in several aspects, we observed important differences between fetal and adult EPDCs. Fetal and adult cells display equal migration abilities in their epithelial state. However, while TGFβ stimulation enhanced adult EPDC migration, it resulted in a reduced migration in fetal EPDCs. Matrigel assays revealed the ability of adult EPDCs to form tube-like structures, which was absent in fetal cells. Furthermore, we observed that fetal cells progress through EMT faster and undergo spontaneous EMT when TGFβ signaling is not suppressed, indicating that fetal EPDCs more rapidly respond to environmental changes. Conclusions Our data suggest that fetal and adult EPDCs are in a different state of activation and that their phenotypic plasticity is determined by this activation state. This culture system allows us to establish the cues that determine epicardial activation, behavior, and plasticity and thereby optimize the adult response post-injury. |
| Databáze: | OpenAIRE |
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