Electrospun Scaffolds as Cell Culture Substrates for the Cultivation of an In Vitro Blood–Brain Barrier Model Using Human Induced Pluripotent Stem Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials and Chemicals
2.2. Preparation of Solutions and Electrospinning
2.3. Crosslinking of Electrospun Fibers
2.4. Scanning Electron Microscopy and Analysis of Fiber Mats
2.5. Confocal Raman Microscopy
2.6. Attenuated Total Reflection-Infrared (ATR-IR) Spectroscopy
2.7. Thickness of Fiber Scaffolds
2.8. Tensile Testing
2.9. Surface Wettability
2.10. Quantification of Released Sodium Fluorescein from Electrospun Fibers
2.11. Preparation of Electrospun Fibers for Cell Cultivation
2.12. Cultivation of Immortalized Human Brain Microvascular Endothelial Cells
2.13. Cultivation and Differentiation of Human Induced Pluripotent Stem Cells
2.14. Evaluation of Cell Viability
2.15. Immunofluorescence Staining
2.16. Impedance Spectroscopy
2.17. Determination of Permeability Coefficients
2.18. Statistical Analysis
3. Results and Discussion
3.1. Fiber Fabrication and Crosslinking
3.2. Physical Characterization of Electrospun Scaffolds
3.3. Cultivation of the Blood–Brain Barrier on Electrospun Scaffolds
3.4. Immunofluorescence Imaging of Tight Junctions
3.5. Evaluation of Barrier Properties
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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GA Concentration (v/v)/ Crosslinking Time | 5% | 15% | 25% |
---|---|---|---|
60 min | X | (✓) | (✓) |
90 min | X | (✓) | ✓ |
120 min | X | (✓) | ✓ |
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Rohde, F.; Danz, K.; Jung, N.; Wagner, S.; Windbergs, M. Electrospun Scaffolds as Cell Culture Substrates for the Cultivation of an In Vitro Blood–Brain Barrier Model Using Human Induced Pluripotent Stem Cells. Pharmaceutics 2022, 14, 1308. https://doi.org/10.3390/pharmaceutics14061308
Rohde F, Danz K, Jung N, Wagner S, Windbergs M. Electrospun Scaffolds as Cell Culture Substrates for the Cultivation of an In Vitro Blood–Brain Barrier Model Using Human Induced Pluripotent Stem Cells. Pharmaceutics. 2022; 14(6):1308. https://doi.org/10.3390/pharmaceutics14061308
Chicago/Turabian StyleRohde, Felix, Karin Danz, Nathalie Jung, Sylvia Wagner, and Maike Windbergs. 2022. "Electrospun Scaffolds as Cell Culture Substrates for the Cultivation of an In Vitro Blood–Brain Barrier Model Using Human Induced Pluripotent Stem Cells" Pharmaceutics 14, no. 6: 1308. https://doi.org/10.3390/pharmaceutics14061308
APA StyleRohde, F., Danz, K., Jung, N., Wagner, S., & Windbergs, M. (2022). Electrospun Scaffolds as Cell Culture Substrates for the Cultivation of an In Vitro Blood–Brain Barrier Model Using Human Induced Pluripotent Stem Cells. Pharmaceutics, 14(6), 1308. https://doi.org/10.3390/pharmaceutics14061308