Epidermal Cells: Methods and Protocols, Vol. 585 Book

Preface v

Contributors xi

1 Isolation of Adult Mouse Stem Keratinocytes Using Magnetic Cell Sorting (MACS) Corina Lorz Carmen Segrettes Marina Garín Jesús M. Paramio 1

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 Michèle T. Martin 13

3 Growth and Stratification of Epithelial Cells in Minimal Culture Conditions Federica Riva Andrea Casasco Marco Casasco Alberto Calligaro Antonia Icaro Comaglia 25

4 Matched Cultures of Keratinocytes and Fibroblasts Derived from Normal and NER-Deficient Mouse Models Alex Pines Claude Backendorf 45

5 Establishment of Spontaneously Immortalized Keratinocyte Lines from Wild-Type and Mutant Mice Julia Reichelt Ingo Haase 59

6 Study of Epidermal Differentiation in Human Keratinocytes Cultured in Autocrine Conditions Frédéric Minner Françoise Herphelin Yves Poumay 71

7 Directed Differentiation of Human Embryonic Stem Cells to Epidermal Progenitors Christian M. Metallo Lin Ji Juan J. de Pablo Sean P. Palecek 83

8 Expression and Analysis of Exogenous Proteins in Epidermal Cells Lina Dagnino Ernest Ho Wing Y. Chang 93

9 Using siRNA Knockdown in HaCaT Cells to Study Transcriptional Control of Epidermal Proliferation Potential Julie Wells Xing Dai 107

10 RNA Interference in Keratinocytes and an Organotypic Model of Human Epidermis Cory L. Simpson Shin-ichiro Kojima Spiro Getsios 127

11 Scanning for Transcription Factor Binding by a Variant EMSA Igor Gurevich Carmen Zhang Brian J. Aneskievich 147

12 Chromatin Immunoprecipitation for Identifying Transcription FactorTargets in Keratinocytes Kori Ortt Satrajit Sinha 159

13 Gene Expression Profiling of Mouse Epidermal Keratinocytes Ramón García-Escudero Jesús M. Paramio 171

14 Analysis of Tissue-Specific Gene Expression Using Laser Capture Microdissection Martin Ruetze Katrin Lochner Stefan Gallinat Anja Knott 183

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 Miroslav Blumenberg 193

16 Molecular Profiling of the Epidermis: A Proteomics Approach Jianjun Shen Susan M. Fischer 225

17 Detection of Gene Expression in Embryonic Tissues and Stratified Epidermis by In Situ Hybridization Maria I. Morasso 253

18 Embryonic Mammary Anlagen Analysis Using Immunolabelling of Whole Mounts Heena Panchal Olivia Wansbury Beatrice A. Howard 261

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 Matthew J. Hardman 271

20 Tetracycline-Regulated Gene Expression in Transgenic Mouse Epidermis Rose-Anne Romano Satrajit Sinha 287

21 A Versatile Murine 3D Organotypic Model to Evaluate Aspects of Wound Healing and Epidermal Organization Eve Kandyba Malcolm Hodgins Patricia Martin 303

22 Optical and Biochemical Dissection of Connexin and Disease-Linked Connexin Mutants in 3D Organotypic Epidermis Stéphanie Langlois Jared M. Churko Dale W. Laird 313

23 Cytokine Release in Tissue-Engineered Epidermal Equivalents-After Prolonged Mechanical Loading Lisette H. Comelissen Debbie Bronneberg Susan Gibbs, Carlijn V.C. Bouten Cees W.J. Oomens 335

24 Three-Dimensional Human Tissue Models of Wounded Skin Christophe Egles Jonathan A. Garlick Tulia Shamis 345

25 In Vivo Transplantation of Genetically Modified Mouse Embryonic Epidermis Ana Belén Martinez-Cruz Clotilde Costa Cristina Saiz Jesús M. Paramio Mirentxu Santos 361

26 A Transplant Model for Human Epidermal Skin Regeneration Sophie Paquet-Fifield Richard P. Redvers Normand Pouliot Pritinder Kaur 369

27 Identification of Epithelial Stem Cells in Vivo and In Vitro Using Keratin 19 and BrdU Danielle Larouche Amélie Lavoie Claudie Paquet Carolyne Simard-Bisson Lucie Germain 383

28 Isolation and Culture of Hair Follicle Pluripotent Stem (hfPS) Cells and Their Use for Nerve and Spinal Cord Regeneration Yasuyuki Amoh Robert M. Hoffman 401

29 Limiting Dilution Analysis of Murine Epidermal Stem Cells Using an In Vivo Regeneration Assays Lauren R. Strachan Ruby Gbadtally 421

Subject Index 433