Epidermal Cells: Methods and ProtocolsKursad Turksen Springer Science & Business Media, 03.02.2008 - 480 Seiten A collection of cutting-edge methods to analyze and manipulate epidermal cellprecursors and mature epidermal cells. These protocols cover different methods and models for culturing epidermal cells, for enriching very early epidermal progenitors, and 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. These readily reproducible techniques broaden our understanding of the biology of epidermal cells and of their utility in normal tissue homeostasis and regenerative medicine applications. |
Inhalt
| 3 | |
Richard P Redvers and Pritinder Kaur 3 Keratinocyte Culture in the Absence of Substrate Attachment | 15 |
Monika Jost and Ulrich Rodeck 4 Application of Genetically Modified Feeder Cells for Culture | 23 |
Takashi Kameda and Toshihiro Sugiyama 5 Organ Culture of Developing Mouse Skin and Its Application | 29 |
for Molecular Mechanistic Studies of Morphogenesis | 39 |
of Cultured Keratinocytes In Vitro and In Vivo | 47 |
Alexander Margulis Weitian Zhang and Jonathan A Garlick | 61 |
In Vivo Labeling and Analysis of Epidermal Stem Cells | 73 |
Characterization of Epithelial Cells in the Hair Follicle | 209 |
Cell Envelopes and Antigen Retrieval | 223 |
Flow Cytometry | 239 |
Fluorimetric DNA Assay of Cell Number | 251 |
Flores and Brian J Aneskievich | 263 |
TetracyclineRegulated Gene Expression in Epidermal Keratinocytes | 273 |
Gene Targeting by Oligonucleotides in Keratinocytes | 287 |
David F Fischer and Claude Backendorf 32 Stable Integration of Large PAC Constructs in Keratinocytes | 303 |
FACS Enrichment of Human Keratinocyte Stem Cells | 87 |
Isolation Characterization and Culture of Epithelial Stem Cells | 97 |
Danielle Larouche Cindy Hayward Kristine Cuffley | 103 |
Immunolocalization in the Epidermis | 113 |
Epidermal Cell Analysis by RTPCR | 121 |
WholeMount Assays for Gene Induction and Barrier Formation | 127 |
Ashley E Webb and David Kimelman | 137 |
Analysis of E2F Factors During Epidermal Differentiation | 147 |
in the Epidermis | 157 |
Apoptosis in the Epidermis | 171 |
Analysis of Connexin 43 Expression on Keratinocytes | 193 |
MMP9 and TIMP1 Assays in Keratinocyte Cultures | 201 |
Sarah H Williams and Alain Hovnanian 33 Targeted Somatic Mutagenesis in the Mouse Epidermis | 315 |
Daniel Metzger Mei Li and Pierre Chambon | 329 |
Methods to Study ProteinProtein Interactions | 341 |
Isolation of Recombinant PhageDisplayed Antibodies | 359 |
Analysis of TissueSpecific DNA Methylation During Development | 371 |
Serial Analysis of Gene Expression in Human Keratinocytes | 383 |
Methods for Gene Expression Profiling in Dermatology Research Using | 399 |
TwoPhoton Fluorescence Imaging and Reactive Oxygen Species | 413 |
In Vivo Transplantation of Engineered Human Skin | 425 |
Gene and Stem Cell Therapy of the Hair Follicle | 437 |
| 449 | |
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acid acrylamide aliquots analysis antibody apoptosis approx assay basal Biol Biology blotting buffer cDNA cell lines centrifuge clones collagen concentration containing cover slip culture medium dermal Dermatol dermis detection dH2O differentiation digestion diluted dissolve EDTA electrophoresis enzyme epidermal Epidermal Cells epidermal keratinocytes epithelial ethanol fibroblasts filter flow cytometry fluorescence forceps gene expression genomic gently graft growth hair follicle human keratinocytes hybridization Incubate interactions Invitrogen keratin keratinocytes labeling layer membrane mg/mL mice microarray Molecular mouse NaCl Note organotypic cultures overnight pellet phage pipet plasmid plate Prepare probe protein Qiagen reaction reagents Remove resuspend Rinse Roche Applied Sciences room temperature sample secondary antibody sections serum Sigma skin slides sodium staining stem cells sterile stock solution store at 20°C Subheading supernatant target tissue transfection transfer transgenic Tris-HCl trypsin tube vector vitro vivo Wash µg/mL