细胞功能测定

Although recent molecular biology advances provide a very effective way in determining localized gene expression, the visualization of the expression by in situ hybridization of whole-mount embryos or paraffin-embedded tissue sections continues to be an excellent method to det ...

Mouse skin has been used extensively as a model system to study the development of cancer. Recent emphasis has been focused on elucidating the molecular mechanisms by which chemical carcinogens or tumor promoters cause the growth of cutaneous malignancies. In this regard, many transgenic ...

The skin as a surface organ is uniquely accessible for whole embryo/foetal analyses of developmental changes, such as gene induction, protein expression, formation of epidermal-derived appendages such as hair follicles and formation of the protective barrier. Such analyses have em ...

The functional and molecular characterization of mouse epidermal stem cells (SC) requires in many circumstances arduous technologies and elaboration of complicated transgenic mouse models. To avoid these difficulties, we have developed a simplified protocol to obtain prima ...

Three-dimensional (3D) organotypic models are increasingly being used to study aspects of epidermal organisation and cutaneous wound-healing events. These are largely dependent on laborious histological analysis and immunohistochemical approaches. Here we outline a me ...

The ability to specifically manipulate gene expression in vivo using mouse models has been one of the most important advances in understanding gene function over the last few decades. Methods used to control gene activities in the mouse include gene targeting and transgenic approaches. Wh ...

The epidermis is a complex tissue composed principally of differentiated keratinocytes that form a keratinized stratified squamous epithelium. The gap junction proteins, connexins (Cx), are differentially expressed throughout the stratified layers of the epidermis and the ...

Prolonged mechanical loading of soft tissues may result in degeneration of these tissues, resulting in formation of pressure ulcers. The risk assessment of individuals might be improved by including measurements of the tissue response to mechanical loading. Cytokines, which are rel ...

Situations in which epidermal mutant mice display early lethality after birth are rather frequent. This condition precludes any kind of analysis of adult or even newborn mice tissues. We propose the in vivo embryonic skin transplantation as an alternative to solve this problem. This method ...

Human skin equivalents (HSEs) are in vitro tissues in which a fully differentiated, stratified squamous epithelium is grown at an air–liquid interface on a Type I collagen gel harboring human dermal fibroblasts. HSEs now provide experimental human tissue models to study factors that dire ...

Progress in the identification of skin stem cells and the improvement of culture methods open the possibility to use stem cells in regenerative medicine. Based on their quiescent nature, the development of label retention assays allowed the localization of skin stem cells in the bulge region ...

This protocol describes a technically simple in vivo assay of long-term skin regeneration in human skin, providing a reliable method for epidermal tissue reconstitution using small numbers of several types of epithelial cells, from epithelial cell lines to primary epithelial stem ce ...

The hair follicle is dynamic, cycling between growth (anagen), regression (catagen), and resting (telogen) phases throughout life. We have demonstrated that nestin-expressing hair follicle stem cells give rise to follicle structures during early anagen or growth phase of the hair fol ...

The basal layer of human interfollicular epidermis is thought to contain a minor compartment of quiescent or slowly cycling epithelial stem cells. These primitive keratinocytes give rise to the progenitors, which are the proliferating keratinocytes and which can be defined as early to l ...

Epidermal stem cells are of major importance for tissue homeostasis, wound repair, tumor initiation, and gene therapy. Here we describe an in vivo regeneration assay to test for the ability of keratinocyte progenitors to maintain an epidermis over the long-term in vivo. Limiting dilution an ...

Biological risk management is required in modern tissue engineering. Particular attention should be paid to the culture medium and the scaffold used. In this perspective, it is important to define minimal culture conditions which allow proper growth and differentiation of epitheli ...

The uppermost layer of our skin, the epidermis, is formed largely of keratinocytes which constitute the skin’s major barrier function and the first line of defence against environmental physical, chemical and biological agents. The subsequent layer, the dermis, which is mainly formed by f ...

A considerable number of transgenic or knockout mice in which epidermal keratinocytes have been targeted die shortly after birth due to barrier defects. In this case, recovery and cultivation of keratinocytes from these animals provide an opportunity for in vitro studies. Working with i ...

Human embryonic stem (hES) cells can differentiate into virtually all somatic cell types. In order to incorporate these derivatives into scientific or clinical applications, efficient methods of directing hES cell differentiation to pure subpopulations are required. Here we des ...

This chapter deals with protocols to set up human keratinocyte cultures in serum-free conditions and lead them to autocrine autonomously growing conditions. These conditions have proven adequate for studies of epidermal differentiation by measurements of the expression of typi ...