Skin Tissue Engineering

This second 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. Chapters describe the isolation, culture of diverse skin cells, skin models, and managing these cells within co-culture systems. Written in the highly successful Methods in Molecular Biology series format, the chapters include brief introductions to the material, lists of necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and a Notes section which highlights tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Skin Tissue Engineering: Methods and Protocols, Second Edition aims to be comprehensive guide for researchers in the field.

Promotion of Human Epidermal Keratinocyte Expansion in Feeder Cell Co-culture. - Isolation and Culture of Epidermal Melanocytes. - Initiating Separate Cultures of Keratinocytes and Melanocytes from a Single Biopsy. - Long-term Expansion of Mouse Primary Epidermal Keratinocytes Using a TGF-beta Signaling Inhibitor. - Isolation and Culture of Hair Follicle Dermal Sheath Mesenchymal Stromal Cells. - Isolation and Culture of Human Dermal Fibroblasts. - Isolation and Culture of Human Dermal Microvascular Endothelial Cells Jennifer Bourland. - Isolation of stromal vascular fraction by fractionation of adipose tissue. - Engineering a multilayered skin equivalent: the importance of endogenous extracellular matrix and model flexibility for nuanced and accurate skin studies. - Engineering a Multilayered Skin Substitute with Keratinocytes, Fibroblasts, Adipose Derived Stem Cells and Adipocytes. - Isolation and Culture of Epidermolysis Bullosa Cells and Organotypic Skin Models. - Engineering a pigmented skin equivalent that is responsive to external stimuli. - Bioprinting a tri-layered skin analogue. - Transdifferentiation of human dermis derived fibroblasts into iNeurons and cultivation in a collagen type 1 hydroge. - Deep proteome profiling of primary skin fibroblasts. - Characterization of human skin derived cells by Raman micro-spectroscopy. - Standard Preparation Protocol of Human Skin and Skin Substitute Samples for Transmission Electron Microscopy. - Methods for assessing scaffold vascularization with human endothelial cells. - Histological and histochemical methods for the evaluation of tissue engineered skin substitutes. - Pig Model to Test Tissue engineered Skin. - Transplantation of Autologous Dermo-Epidermal Skin Substitutes in a Pig Model. - Ex vivo human skin model to study immediate and early local responses to external stimuli. - A Method to Evaluate Skin and Skin Substitute Mechanical Properties Using a Suction Device. - GMP compliant production of therapeutic components of autologous adipose tissue. - Quality controls of autologous skin products according to European GMP guidelines. - Standardized outcome measures for the clinical application of tissue engineered products.