Abstract
Isolation and culture of keratinocytes from patients with different types of epidermolysis bullosa are sometimes challenging, because of the inherent adhesion defects of these cells. We routinely employ a well-established protocol for in vitro culture of these cells from small skin samples remaining after diagnostic procedures are performed. Keratinocytes and fibroblast can be used for downstream expression and functional studies or for construction of in vitro organotypic cocultures. These cells maintain main common characteristics of spreading, adhesion, migration, and survival, which depend on the underlying molecular defect.
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Abbreviations
- DMEM:
-
Dulbecco’s modified Eagle medium
- EB:
-
Epidermolysis bullosa
- Fb:
-
Fibroblasts
- FCS:
-
Fetal calf serum
- PBS:
-
Phosphate-buffered saline
References
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Acknowledgments
The authors would like to thank current and former members of the laboratory. A special thanks is said to Juna Leppert team members who have contributed to adapt the protocols to obtain this for her valuable suggestions in the manuscript.
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He, Y., Has, C. (2019). Isolation and Culture of Epidermolysis Bullosa Cells and Organotypic Skin Models. In: Böttcher-Haberzeth, S., Biedermann, T. (eds) Skin Tissue Engineering. Methods in Molecular Biology, vol 1993. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9473-1_14
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DOI: https://doi.org/10.1007/978-1-4939-9473-1_14
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Publisher Name: Humana, New York, NY
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