This second edition volume expands on the previous edition with a discussion on the latest advancements in laboratory techniques used in dermatological research. The chapters in this book cover topics such as methodologies for establishing 2D and 3D skin models; assessing DNA damage; genome editing using CRISPR/Cas systems; analyzing extracellular vesicles involved in intercellular communication; and detecting microflora and skin permeability barrier. This book also presents experimental strategies for modeling and modulating cutaneous pathologies, including wound healing and its associated microbiome, skin cancer, and hair follicle disorders. Written in the highly successful Methods in Molecular Biology series format, 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 thorough, Molecular Dermatology: Methods and Protocols, Second Editon is a valuable resource for researchers working in both fundamental and applied dermatological science.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key implementation advice from the experts

Inhalt

Isolation of Epidermal Keratinocytes from Human Skin: The Scratch-Wound Assay for Assessment of Epidermal Keratinocyte Migration.- Isolation of Different Dermal Fibroblast Populations from the Skin and the Hair Follicle.- Isolation of Epidermal and Hair Follicle Melanocytes.- Validated Step-By-Step Protocols for Human Keratinocyte Isolation and the Construction of Organotypic 3D Epidermal Models.- Human Epidermal Equivalents on Non-Woven Scaffolds: A Model of Tissue Formation in an Artificial Environment.- Purification of Extracellular Vesicles Secreted by Skin Fibroblasts and Keratinocytes.- Genetic Engineering of Human Keratinocytes using CRISPR/Cas9 Ribonucleoprotein Complexes or Modified Cas9-Encoding mRNAs.- Imaging Approaches to DNA Damage Induction and Quantification.- Identification of Long Non-Coding RNA by In Situ Hybridization Approaches.- Detection of MicroRNAs in Skin by In Situ Hybridization.- 3D-FISH Analysis of the Spatial Genome Organization in Mammalian Tissue in Situ.- Methods to Investigate the Epidermal Permeability Barrier and the Light Irradiation Barrier In Vitro and Ex Vivo.- A Protocol to Prepare and Stain Dorsal Skin Whole Mounts from Mice.- Isolation of Mouse Hair Matrix Keratinocytes for Multi-Omics Analysis.- Methods to Study Human Hair Follicle Growth Ex Vivo: Human Microdissected Hair Follicle and Human Full Thickness Skin Organ Culture.- Generation of Hair Follicle Germs In Vitro Using Human Postnatal Skin Cells.- In Situ Laser Capture Microdissection for Detection of Components of the Hair Follicle and Scalp Microbiome.- Viable vs. Non-Viable Microbiota Evaluation of the Hair Follicle and Scalp Microbiome.- Non-Surgical Induction of Alopecia Areata in C3H/HeJ Mice via Adoptive Transfer of Cultured Lymphoid Cells.- Collapse and Restoration of Hair Follicle Immune Privilege Ex Vivo: A Model for Alopecia Areata.- Experimentally Induced Epithelial-Mesenchymal Transition of Human Hair Follicle Stem Cells as a Model of Scarring Alopecia Ex Vivo.- Method for Human Sweat Gland Isolation from the Scalp and Axilla by Means of the Micro-Punch FUE Technique and Vital Dyes.- Assessment of Acute Wound Healing using Excisional Mouse Tail Skin Wound Models.- Skin and Wound Infection Models to Test Host-Pathogen Interactions and Evaluate Topical Antimicrobial Treatments.- Human Wound Healing Ex Vivo Model with Focus on Molecular Markers.- The Evaluation of Fibrotic Alterations in Regenerated Murine and Human Xenografted Skin.- Method to Study Skin Cancer: Two-Stage Chemically Induced Carcinogenesis in Mouse Skin.- Ultraviolet Irradiation of Skin Cells.- Method for Investigation of Photobiological Effects of Light on Human Skin Cells In Vitro Using Visible and Near-Infrared Light.