细胞技术

Proteomics is the term used for the large-scale analysis of proteins in biological fluids or cells by biochemical methods. Two approaches are used for proteomics analysis: two-dimensional polyacrylamide gel electrophoresis (2D-PA GE) and a mass-spectrometry-based approach, su ...

Successful gene profiling studies involve careful experimental design, use of sensitive and accurate technologies, and statistically valid analysis of experimental results. In this chapter we describe our approach to the profiling of pancreatic adenocarcinoma to illustra ...

Serial analysis of gene expression (SAGE) is a molecular biology technique that was developed to measure the global gene expression levels. It has been applied successfully to characterize transcriptomes, compare the transcript levels between normal and diseased tissues, and unco ...

The recognition of a homozygous deletion of genetic material in a tumor genome has been instrumental in several tumor suppressor gene searches. The representational difference analysis (RDA) allows one to identify homozygous deletions even from among the high background of allelic l ...

Digital single-nucleotide polymorphism (SNP) analysis is developed to amplify a single template from a pool of DNA samples, thereby generating the amplicons that are homogeneous in sequence. Different fluorophores are then applied as probes to detect and discriminate different al ...

Many tumor suppressor genes (such as p16, Rb, VHL, E-cadherin, and hMLH1) that are silenced by mutation are also inactivated by gene silencing through DNA methylation. Characterization of genes hypermethylated in human cancers but not in normal tissues not only provides insights into canc ...

Some populations of the epithelial cells from the duct and ductular network of the mammalian pancreas have been isolated and maintained in vitro for up to 3 mo. These cells express many of the surface factors that are unique to them in vivo. They also retain significant drug-and carcinogen-metabol ...

Xenotransplantation (xenografting) of primary cancers or cancer cell lines into immunodeficient mice is a commonly used technique to assess tumor growth in response to a variety of experimental agents. When primary pancreatic cancers are xenografted, cancer cells proliferate in ...

The tissue microarray (TMA) of Kononen et al. is an extension of an idea originally developed by Battifora and consists of an array of cylindrical cores of paraffin-embedded tissue that are removed from preexisting “donor” paraffin blocks. The donor block is a standard tissue block that may be from ...

There are many types of pancreatic neoplasms. Pathologic examination, which includes both routine (e.g., hematoxylin-and-eosin staining) and ancillary (e.g., immunohistochemistry and in situ hybridization) techniques, is essential in correctly typing a pancreatic neopla ...

Isolation of well-preserved pure cell populations is a prerequisite for sound studies of the molecular basis of pancreatic malignancy and other biological phenomena. This chapter reviews current methods for obtaining anatomically specific signals from molecules isolated fr ...

Histologically distinct noninvasive precursor lesions have been recognized in the pancreas for close to a century. The recent development of a consistent reproducible nomenclature and classification system for these lesions has been a major advance in the study of these noninvasi ...

Collagen-induced arthritis (CIA) is an animal model of rheumatoid arthritis (RA) that is widely used to address questions of disease pathogenesis and to validate therapeutic targets. Arthritis is normally induced in mice or rats by immunization with autologous or heterologous type II c ...

With the exponential increase in the number of genes identified by various genome projects, it has become imperative that efficient methods be developed for deciphering gene function. Genetically engineered strains of mice are now critical research tools for basic biomedical resea ...

Tumor necrosis factor α (TNFα) appears to be the main mediator causing the systemic manifestations of sepsis and septic shock associated with edema (1,2). In vitro it augments movement of permeability tracers across endothelial cell monolayers (3,4). Herein we describe a method which perm ...

In most cellular systems tumor necrosis factor (TNF) induces apoptotic cell death. However, in some particular cell lines, such as the L929sA fibrosarcoma, TNF induces necrotic cell death. This effect is not the result of an inability to die apoptotically, because triggering of Fas in L929sAh ...

The protocols for immunohistochemical localization of tumor necrosis factor α (TNFα), TNFRp55, and TNFRp75 in central nervous system (CNS) slices from laboratory animals are described. The procedure requires intracardiac perfusion of animals followed by fixation and cryostat ...

Polymorphisms in the promoter of the tumor necrosis factor alpha (TNFα) gene have been reported to affect the transcription rate and the release of this cytokine. Among the best studied, the −238 and −308 polymorphisms have been associated with an increased transcriptional activity and TN ...

Tumor necrosis factor (TNF) is a pleiotropic cytokine with a wide range of biological activities including cytotoxicity, immune-cell proliferation, and mediation of inflammatory responses. Mutational analysis of mature TNF has been facilitated by the high expression levels th ...

The finding that the two tumor necrosis factor receptors (TNFR) exist in soluble form in various body fluids not only has substantiated the paradigm of naturally existing soluble cytokine receptors but also has represented a milestone on the road to the biochemical and biological charact ...