Skin Stem Cells Methods and Protocols

This updated edition explores techniques in identifying and characterizing various skin stem cells and generating skin-like structures in vitro. The goal of the book is to present exciting advances in skin biology in an effort to provide a better understanding of skin and, eventually, to allow us to regenerate skin with its original form and function. 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 and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and up-to-date, Skin Stem Cells: Methods and Protocols, Third Edition serves as a valuable assemblage of protocols for researchers already working in the field and for those who now wish to newly embark on studies of skin stem cells.

Isolation, Characterization, and Utilization of Human Skin Basal and Supra-Basal Epidermal Stem Cells.- Isolation of Human Progenitor Epidermal Cells on Collagen Type IV and Analysis of Their Markers with Flow Cytometry and PCR Methods.- Approaches to Study Wound-Induced Hair Neogenesis (WIHN).- Characterization of the Newborn Epidermis and Adult Hair Follicles Using Whole-Mount Immunofluorescent Staining of Mouse Dorsal Skin.- Preparation and Use of shRNA for Knocking Down Specific Genes.- Efficient Generation of CRISPR/Cas9-Mediated Knockout Human Primary Keratinocytes by Electroporation.- Application of CRISPR/Cas9 Pooled Screening for a Non-Immediate Readout Model.- Contribution of Artificial Intelligence to the Identification of Protein-Protein Interactions: A Case Study on PAR-3 and Its Partner Adapter Molecule Crk.- Site-Directed Mutagenesis to Mutate Multiple Residues in a Single Reaction.- Analysis of Proliferation, Apoptosis, and Motility in Mouse Embryonic Melanocytic Precursor Cells.- Protocol for Testing Human Melanoma Exosomes in Shifting Healthy Phenotype of Human Dermal Cells.- Incorporating NK Cells in a Three-Dimensional Organotypic Culture System for Human Skin Stem Cells: Modeling Skin Diseases and Immune Cell Interplay.- Standardized GMP-Compliant Scalable 3D-Bioprocessing of Epidermal Stem Cells for Diabetic Foot Ulcers.- Differentiation and Validation of Human Induced Pluripotent Stem Cells into a Keratinocyte, as Therapeutic Option in Vitiligo.- GMP-Based Isolation of Full-Term Human Placenta-Derived NK Cells for CAR-NK Cell Therapy in Malignant Melanoma.- Isolation and Phenotypic Characterization of Tumor-Infiltrating NK Cells in Skin Carcinoma.- Preparation and Validation of Zebrafish Psoriasis Model to Investigate the Therapeutic Effects of Stem Cells.- Flow Cytometric and Immunohistochemical Follow-Up of Spermatogonial Lineage Commitment.- Development of CAR-NK Cells Targeting cSCC-Specific Antigens for Precision Immunotherapy.