细胞功能测定

Polycations have been used as nonviral gene carriers because the polycations and DNAs form stable complexes in a noncovalent manner (1–3). The polycations, e.g., poly(L-lysine) (PLL), are reported to be conjugated with several ligands for targeted gene delivery (4–8). The physicochemical ...

The tetracycline-regulated expression system developed by Gossen and Bujard is a powerful genetic tool that permits the expression of any gene construct introduced into either cultured cells or transgenic animals to be precisely controlled. It involves two components, a regulato ...

Protocols to study the regulation of a conserved multigene family (SPRR genes) during calcium-induced differentiation of cultured normal human keratinocytes (NHKs) are provided. Transfection of promoter-reporter (CAT or luciferase) constructs, combined with promoter tr ...

Oligonucleotide-directed gene alteration produces a targeted deoxyribonucleic acid (DNA) sequence change in the genome of mammalian cells at low frequency that is only detectable by highly sensitive methods. To measure the low frequency, we have established an assay using the mutant ...

Transfer of P1-derived artificial chromosome (PAC) deoxyribonucleic acid (DNA) into keratinocytes is an extremely important technique that enables functional studies of keratinocyte-specific genes to be performed and genomic gene therapy for inherited and acquired disea ...

Protein-protein interactions are the underpinnings of a vast number of cellular processes. In recent years, the convergence of biochemistry, cellular, and molecular biology has made available a number of powerful techniques for studying such interactions. These techniques vary in ...

The efficient introduction of somatic mutations in a given gene at a given time and in specific cell types of the skin will greatly facilitate the studies of a number of genes expressed in the skin and the production of animal models for skin diseases. We describe here strategies and techniques to create ...

Advances in our understanding of biology at the molecular level are very much driven by improvements in the scientist’s tool box. Such improvements may not only be an introduction of new techniques like polymerase chain reaction, but as much an increment of for example the sensitivity of existi ...

The demand for large-scale gene expression analysis tools is on the rise now that several genomes have been sequenced. One of these tools, serial analysis of gene expression (SAGE), allows the qualitative as well as quantitative analysis of a large number of genes in a defined tissue or culture model. ...

The establishment of a cell identification method more accurate than the conventional morphological and cell-type-specific marker analyses has been desired. DNA methylation is related to the gene activity including gene-silencing and is a key mechanism of epigenetics underling ...

Here we present methods of gene expression profiling using nylon filter deoxyribonucleic acid (DNA) microarrays and radiolabeled and nonradiolabeled hybridization probes. DermArray� nylon filter DNA microarrays were designed specifically for use in dermatology resear ...

Two-photon fluorescence microscopy is used to detect ultraviolet-induced reactive oxygen species (ROS) in the epidermis and the dermis of ex vivo human skin and skin equivalents. Skin is incubated with the nonfluorescent ROS probe dihydrorhodamine, which reacts with ROS such as singl ...

The skin is an important target for gene transfer because of its easy accessibility. Recently, we demonstrated that an in vivo electroporation approach could be applied to plasmid deoxyribonucleic acid delivery in rat skin of the abdominal area. Plate-and-fork type electrodes were effe ...

Tissue engineering approaches have enabled the development of methodologies that allow long-term, in vivo studies in epidermal biology. This has been accomplished through in vivo transplantation of human epidermal cells fabricated as three-dimensional, skin-equivalents in ...

The hair follicle is a highly complex appendage of the skin containing a multiplicity of cell types. The follicle undergoes constant cycling through the life of the organism including growth and resorption with growth dependent on specific stem cells. The targeting of the follicle by genes and ...

Mesenchymal cells have been cultured successfully for many years owing to the ease of their culturability. With the advent of more complex growth media, and some changes to methods, it has become possible to culture epithelial cell types from a wide variety of rodent and human tissue sources succ ...

Cell culture has become an integral part of the daily routine of most oncology laboratories. It has enabled researchers to investigate a wide range of cellular parameters in a defined system in which the experimental conditions can be controlled and repeated. Although the manufacturers of t ...

Because of its cellular complexity and regional heterogeneity, the mammalian central nervous system is not easily amenable for experimental analysis. The study of the developing brain becomes even more complicated because of the differential growth rates of different parts of the br ...

The thyroid gland contains two populations of epithelial cells, of quite different embryological origin and function. Only the major component—the follicular cells—will be considered here. The minor C cell population, which forms only a few percent, can be ignored for the purposes of prim ...

With the advent in 1969 of the collagenase-perfusion technique for the high-yield preparation of isolated, differentiated hepatocytes (1), easy establishment of primary cultures of hepatocytes was made possible. Since then, this experimental system has been increasingly used in ...