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This volume provides protocols describing the isolation and culture of diverse cell types stemming from the skin and the use of these cells and cell constructs for wound healing, bioengineering applications, and translational medicine purposes. The book is divided into three sections describing the isolation and culture of diverse skin cells, managing these cells within co-culture systems and skin models, as well as using these skin models in a test setting. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Skin Tissue Engineering: Methods and Protocols serves as a vital aid to basic and clinical researchers such as biologists, physicians, and biomedical engineers working with and being interested in basic science, and clinically and laboratory-applicable translational regenerative medicine.
List of contents
Part 1 : Isolation and culture of skin cells.- Chap 1: Isolation and Culture of Human Keratinocytes.- Chap 2 : Promotion of Human Epidermal Keratinocyte Expansion in Feeder Cell Co-culture.- Chap 3 : Isolation and Culture of Epidermal Melanocytes.- Chap 4: Long-term Expansion of Mouse Primary Epidermal Keratinocytes Using a TGF-beta Signaling Inhibitor.- Chap 5 : Isolation and Culture of Hair Follicle Dermal Sheath Mesenchymal Stromal Cells.- Chap 6 : Isolation and Culture of Human Dermal Fibroblasts.- Chap 7: Isolation and Culture of Human Dermal Microvascular Endothelial Cells.- Chap 8: Isolation of Stromal Vascular Fraction by Fractionation of Adipose Tissue.- Part 2 : Tissue engineering skin and engineered skin models.- Chap 9 : Engineering a Multi-Layered Skin Equivalent: the Importance of Endogenous Extracellular Matrix Maturation to Provide Robustness and Reproducibility.- Chap 10 : Three-dimensional Epidermal Model from Human Hair follicle-derived Keratinocytes.- Chap 11 : Fabrication of a Co-Culture System with Human Sweat Gland Derived Cells and Peripheral Nerve Cells.- Chap 12 : Engineering a Multilayered Skin Substitute with Keratinocytes, Fibroblasts, Adipose Derived Stem Cells and Adipocytes.- Chap 13 : Skin Tissue Engineering Fabrication of Chimeric Hair Follicles for Skin Tissue Engineering.- Chap 14 : Isolation and Culture of Epidermolysis Bullosa Cells and Organotypic Skin Models.- Part 3 : In vitro and vivo models testing skin, skin cells, and tissue engineered skin.- Chap 15 : Effects of the Extracellular Matrix on the Proteome of Primary Skin Fibroblasts.- Chap 16 : Standard Preparation Protocol of Human Skin Samples for Transmission Electron Microscopy.- Chap 17 : Methods for Assessing Scaffold Vascualarization In Vivo.- Chap 18 : Human Reconstructed Skin in a Mouse Model.- Chap 19 : Pig Model to Test Tissue Engineered Skin.- Chap 20 : Transplantation of Autologous Dermo-Epidermal Skin Substitutes in a Pig Model.
