Development of Human Salivary Gland-Like Tissue In Vitro
| Autor: | Matthias Schweinlin, Heike Walles, Simon Lennartz, Rudolf Hagen, Stephan Hackenberg, Gudrun Steusloff, Christian Ginzkey, Marco Metzger, Norbert Kleinsasser, Marc Burghartz, Agmal Scherzad, Kathrin Radeloff |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Pathology medicine.medical_specialty Biomedical Engineering Bioengineering Biology Matrix (biology) Immunofluorescence Biochemistry Salivary Glands Biomaterials 03 medical and health sciences 0302 clinical medicine Tissue engineering medicine Humans Cells Cultured Decellularization Tissue Engineering medicine.diagnostic_test Salivary gland Endothelial Cells Cell Differentiation Epithelial Cells Molecular biology Coculture Techniques Enzyme assay In vitro 030104 developmental biology medicine.anatomical_structure Real-time polymerase chain reaction 030220 oncology & carcinogenesis Amylases Microscopy Electron Scanning biology.protein |
| Zdroj: | Tissue Engineering Part A. 24:301-309 |
| ISSN: | 1937-335X 1937-3341 |
| Popis: | The loss of salivary gland function caused by radiation therapy of the head and neck is a serious condition and it affects a patient's quality of life. The current lack of effective therapies demands new options to be explored. This study tested whether human salivary gland epithelial cells (SGECs) could be successfully cultured on a decellularized porcine gut matrix (SIS-muc) in both mono- and coculture with microvascular endothelial cells (mvECs). By performing immunofluorescence imaging, transmission as well as scanning electron microscopy (SEM), quantitative polymerase chain reaction (qPCR), and an amylase enzyme assay, it was investigated as to what extent the three-dimensional (3D)-cultured cells could maintain their molecular differentiation and the production of working α-amylase (α-AMY) compared with two-dimensional (2D) culture. In both 3D mono- and coculture, SGECs were successfully cultured and formed acinar-like structures. Those findings were confirmed by SEM imaging. Immunofluorescence imaging revealed that 3D-cultured cells expressed α-AMY, Claudin-1 (CL-1), and water channel protein aquaporin-5 (AQP-5). Two-dimensional-cultured cells only were positive for α-AMY. Real time (RT)-qPCR analysis showed that α-AMY relative gene expression was higher in both 3D mono- and coculture than in 2D culture. In α-AMY enzyme assay, cocultured SGECs showed about 25 times increased enzyme activity compared with 2D-cultured cells. In conclusion, the SIS-muc combined with endothelial coculture seems a suitable culture setting for the tissue engineering of functional human salivary gland tissue. |
| Databáze: | OpenAIRE |
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