Main description:

In recent years, our ability to understand and manipulate epidermal cells has increased tremendously, opening significant new possibilities in both basic science research and in regenerative medicine, including would healing and transplantation. Epidermal Cells: Methods and Protocols, Second Edition expands upon the popular first edition by bringing together a panel of experienced basic and clinical researchers to describe in step-by-step detail the powerful methods they have developed and optimized to analyze and manipulate epidermal cell precursors and mature epidermal cells. These protocols cover different methods and models for culturing epidermal cells, for enriching very early epidermal progenitors, as well as for studying epidermal cell commitment and differentiation both in vitro and in vivo. Topics of special interest include the derivation, characterization, and utility of epidermal stem cells, mature epidermal cells and their characterization, and applications in regenerative medicine. The protocols follow the successful Methods in Molecular Biology (TM) series format, providing introductions to their respective topics, lists of the necessary materials and reagents, and notes on troubleshooting and avoiding known pitfalls.

State-of-the-art and highly practical, Epidermal Cells: Methods and Protocols, Second Edition offers experienced and novice investigators alike an invaluable collection of readily reproducible techniques designed to broaden not only our understanding of the biology of epidermal cells, but also their utility in normal tissue homeostasis and regenerative medicine applications.

Contents:

1. Isolation of Adult Mouse Stem Keratinocytes Using Magnetic Cell Sorting (MACS)
Corina Lorz, Carmen Segrelles, Marina Garin, and Jesus M. Paramio

2. Functional Investigations of Keratinocyte Stem Cells and Progenitors at a Single-Cell Level Using Multiparallel Clonal Microcultures
Nicolas O. Fortunel, Pierre Vaigot, Emmanuelle Cadio, and Michele T. Martin

3. Growth and Stratification of Epithelial Cells in Minimal Culture Conditions
Federica Riva, Andrea Casasco, Marco Casasco, Alberto Calligaro, and Antonia Icaro Cornaglia

4. Matched Cultures of Keratinocytes and Fibroblasts Derived from Normal and NER-Deficient Mouse Models
Alex Pines and Claude Backendorf

5. Establishment of Spontaneously Immortalised Keratinocyte Lines from Wild-Type and Mutant Mice
Julia Reichelt and Ingo Haase

6. Study of Epidermal Differentiation in Human Keratinocytes Cultured in Autocrine Conditions
Frederic Minner, Francoise Herphelin, and Yves Poumay

7. Directed Differentiation of Human Embryonic Stem Cells to Epidermal Progenitors
Christian M. Metallo, Lin Ji, Juan J. de Pablo, and Sean P. Palecek

8. Expression and Analysis of Exogenous Proteins in Epidermal Cells
Ernest Ho, Wing Y. Chang, and Lina Dagnino

9. Using siRNA Knock-Down in HaCaT Cells to Study Transcriptional Control of Epidermal Proliferation Potential
Julie Wells and Xing Dai

10. RNA Interference in Keratinocytes and an Organotypic Model of Human Epidermis
Cory L. Simpson, Shin-ichiro Kojima, and Spiro Getsios

11. Scanning for Transcription Factor Binding by a Variant EMSA
Igor Gurevich, Carmen Zhang, and Brian J. Aneskievich

12. Chromatin Immunoprecipitation for Identifying Transcription Factor Targets in Keratinocytes
Kori Ortt and Satrajit Sinha

13.Gene Expression Profiling of Mouse Epidermal Keratinocytes
Ramon Garcia-Escudero and Jesus M. Paramio

14. Analysis of Tissue-Specific Gene Expression Using Laser Capture Microdissection
Martin Ruetze, Katrin Lochner, Stefan Gallinat, and Anja Knott

15. Comprehensive Transcriptional Profiling of Human Epidermis, Reconstituted Epidermal Equivalents and Cultured Keratinocytes, Using DNA Microarray Chips
Ding-Dar Lee, Jiri Zavadil, Marjana Tomic-Canic, and Miroslav Blumenberg

16. Molecular Profiling of the Epidermis: A Proteomics Approach
Jianjun Shen and Susan M. Fischer

17. Detection of Gene Expression in Embryonic Tissues and Stratified Epidermis by In Situ Hybridization
Maria I. Morasso

18. Embryonic Mammary Anlagen Analysis Using Immunolabelling of Whole Mounts
Heena Panchal, Olivia Wansbury, and Beatrice A. Howard

19. Whole-Mount Assays for Gene Induction and Barrier Formation in the Developing Epidermis
Carolyn Byrne, Ariel A. Avilion, Ryan F. O'Shaughnessy, Jonathan C. Welti, and Matthew J. Hardman

20. Tetracycline-Regulated Gene Expression in Transgenic Mouse Epidermis
Rose-Anne Romano and Satrajit Sinha

21. A Versatile Murine 3D Organotypic Model to Evaluate Aspects of Wound Healing and Epidermal Organization
Eve Kandyba, Malcolm Hodgins, and Patricia Martin

22. Optical and Biochemical Dissection of Connexin and Disease-Linked Connexin Mutants in 3D Organotypic Epidermis
Stephanie Langlois, Jared M. Churko, and Dale W. Laird

23. Cytokine Release in Tissue Engineered Epidermal Equivalents after Prolonged Mechanical Loading
Lisette H. Cornelissen*, Debbie Bronneberg, Susan Gibbs, Carlijn V.C. Bouten, and Cees W.J. Oomens

24. Three-Dimensional Human Tissue Models of Wounded Skin
Christophe Egles and Jonathan A.