Molecular Dermatology Methods and Protocols

This volume delivers the most important laboratory techniques that underlie dermatological investigation, including in situ, in vitro, ex vivo, and in vivo models of skin, its appendages (hair follicles, sebaceous and sweat glands), and immune cells. Also included are unique techniques that unravel epigenetic mechanisms in skin and hair follicle cells, such as gene organization in the nucleus and gene expression regulation by the non-coding transcriptome, which are important recent developments in investigative dermatology. 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. 

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.- Isolation and Culture of Human Skin Mast Cells.- Organotypic 3D Skin Models: Human Epidermal Equivalent Cultures from Primary Keratinocytes and Immortalized Keratinocyte Cell Lines.- Purification of Extracellular Microvesicles Secreted by Dermal Fibroblasts.- A Method to Investigate the Epidermal Permeability Barrier In Vitro.- Isolating Dermal Papilla Cells from Human Hair Follicles Using Microdissection and Enzyme Digestion.- Methods to Study Human Hair Follicle Growth Ex Vivo: Human Microdissected Hair Follicle and Human Full Thickness Skin Organ Culture.- Non-Surgical Induction of Alopecia Areata in C3H/HeJ Mice via Adoptive Transfer of Cultured Lymphoid Cells.- Collapse and Restoration of Hair Follicle ImmunePrivilege 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.- Generation of Hair Follicle Germs in Vitro Using Human Postnatal Skin Cells.- Method for Human Eccrine Sweat Gland Isolation from the Scalp by Means of the Micro-Punch Technique and Vital Dyes.- Identification of Long Non-Coding RNA by In Situ Hybridization Approaches.- Detection of MicroRNAs by In Situ Hybridization in Skin.- Chromatin Immunoprecipitation of Low Number of FACS-Purified Epidermal Cells.- 3D-FISH Analysis of the Spatial Genome Organization in Skin Cells In Situ.- Method to Study Skin Cancer: Two-Stage Chemically-Induced Carcinogenesis in Mouse Skin.- Investigation of Skin Wound Healing Using a Mouse Model.- Human Wound Healing Ex Vivo Model with Focus on Molecular Markers.- Method for Investigation of Photobiological Effects of Light on Human Skin Cells Mediated by Low Doses of Light.