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This fully updated edition brings together a new set of protocols to arm epidermal cell biologists with tools and approaches to continue the quest to uncover the intricacies and regulatory mechanisms underlying epidermal cell development and function. The collection takes a detailed view at techniques currently being applied to this developing field. 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.
Comprehensive and cutting-edge, Epidermal Cells: Methods and Protocols, Fourth Edition serves as a valuable reference for experts as well as novices in the epidermal cell field.
List of contents
The Simplest Protocol for Rapid and Long-Term Culture of Primary Epidermal Keratinocytes from Human and Mouse.- Dermal-Epidermal Separation by Heat.- Dermal-Epidermal Separation by Enzyme.- Dermal-Epidermal Separation by Chemical.- Isolation and Characterization of Extracellular Vesicles from Keratinocyte Cultures.- Competitive Repopulation Assay of Long-Term Epidermal Stem Cell Regeneration Potential.- Quantification of Melanosome Transfer Using Immunofluorescence Microscopy and Automated Image Analysis.- Contribution of Immunohistochemistry in Revealing S100A7/JAB1 Colocalization in Psoriatic Epidermal Keratinocyte.- Digital Quantification of Epidermal Protein Expression in Paraffin-Embedded Tissue Using Immunohistochemistry.- Keratinocyte Differentiation by Flow Cytometry.- In Silico and In Vitro Considerations of Keratinocyte Nuclear Receptor Protein Structural Order for Improving Experimental Analysis.- Genetic Modification of Human Primary Keratinoctes by Lentiviral Vectors.- Generation of Knockout Human Primary Keratinocytes by CRISPR/Cas9.- In Vitro Wound Healing Assays to Investigate Epidermal Migration.- Iterative Three-Dimensional Epidermis Bioengineering and Xenografting to Assess Long-Term Regenerative Potential in Human Keratinocyte Precursor Cells.- Generation of a Full-Thickness Human Skin Equivalent on an Immunodeficient Mouse.- Biotin Identification Proteomics in Three-Dimensional Organotypic Human Skin Cultures.- Information and Statistical Analysis Pipeline for High-Throughput RNA Sequencing Data.- Cell-Extracellular Matrix Adhesion Assay.- Methods for Analysis of Keratinocyte Migration.- Scratch Wound Healing Assay.- Photoprotective Activity Assay toward Ultraviolet B in Human Keratinocytes.- Photodynamic Therapy Assay.- A Method to Prepare Claudin-Modulating Recombinant Proteins.- Human Fetal Skin Fibroblast Isolation and Expansion for Clinical Application.- Ciliated Epithelial Cell Differentiation at Air-Liquid Interface Using Commercially Available Culture Media.
