细胞技术

S phase is that period of time in the cell-division cycle during which nuclear chromosomal deoxyribonucleic acid (DNA) is replicated (1,2). The time required for S phase depends on the size of the genome, the organism, and its developmental state. DNA replication requires only 15 to 20 min in budding ye ...

One of the most common uses of flow cytometry is to analyze the cell cycle of mammalian cells. Flow cytometry can measure the deoxyribonucleic acid (DNA) content of individual cells at a rate of several thousand cells per second and thus conveniently reveals the distribution of cells through the ce ...

Understanding the molecular and biochemical basis of cellular functions involved in growth and proliferation requires the investigation of regulatory events that most often occur in a cell cycle phase-dependent fashion. Studies involving cell cycle regulatory mechanisms and ...

When studying cell cycle checkpoints, it is often very useful to have large numbers of cells that are synchronized in various stages of the cell cycle. A variety of methods have been developed to obtain synchronous (or partially synchronous) cells, all of which have some drawbacks. Many cell types t ...

The way cells respond to radiation or chemical exposure that damages deoxyribonucleic acid (DNA) is important because induced lesions left unrepaired, or those that are misrepaired, can lead to mutation, cancer, or lethality. Prokaryotic and eukaryotic cells have evolved mechanisms ...

Omega-3 polyunsaturated fatty acids (PUFAs) such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) display beneficial actions in human diseases. The molecular basis for these actions remains of interest. We recently identified novel mediators generated from ω-3 PUFA ...

This chapter describes in vitro methods for studying the Blood-Brain barrier. These methods include a cell line and isolated brain microvessels. The rat brain endothelial cell line 4 (RBE4) express many properties that are expressed by brain endothelial cells in vivo. Tissue culture meth ...

Lipid rafts are dynamic structures made up of proteins and lipids that float freely within the liquid-disordered bilayer of cellular membranes and have the ability to cluster to form larger, more-ordered platforms. These clustered structures have been identified in all cell types and have ...

Epithelial cells at all mucosal surfaces are potentially apposed to bacteria, particularly in the intestine. It is established that intestinal epithelial cells (IECs) represent an important barrier between lamina propria cells and the potentially harmful lumenal contents. In ad ...

Adhesive interactions between cells regulate tissue integrity as well as the process of inflammatory cell recruitment. Such intercellular interactions are regulated by adhesion receptors and can be homotypic, that is, between cells of the same type, for example, between adjacent en ...

Disrupted epithelial cell junctions are a hallmark of numerous disease processes. The signaling mechanisms regulating barrier function and re-establishment of intact junctions after injury and during development are complex and tightly regulated. We have shown that heterot ...

Models to study molecular, biochemical, and functional responses in vitro generally incorporate an individual cell type or group of cells organized in a random fashion. Normal physiological responses in vivo require that individual cell types be oriented in an organized fashion with t ...

Interactions between the vascular endothelium and polymorphonuclear leukocytes (PMNs) are central to PMN emigration into inflamed tissues, and to neutrophil–endothelial crosstalk pathways that modulate inflammatory responses and vascular barrier function. For exam ...

The interactions between egg and sperm are among the most fascinating in cell biology. These interactions include Cell-Cell adhesion and then membrane fusion between the two gametes. This chapter details the experimental methods used to for gamete culture and in vitro fertilization us ...

Mucosal tissues undergo contraction and relaxation on a continuous basis. In its normal state, the pliable intestinal tract is characterized by a rhythmic pattern of contractions controlled by its intrinsic neuronal innvervation. In chronic inflammatory diseases such as Crohn’s d ...

An essential requirement for adequate organ performance is the formation of permeability barriers that separate and maintain compartments of distinctive structure. The endothelial cell lining of the vasculature defines a semipermeable barrier between the blood and the inter ...

The preparation of mammalian mitochondrial ribosomes in sufficient quantities for biochemical studies is best done beginning with whole tissue rather than from cells in culture. This issue arises because of the low abundance of these ribosomes in cells, making their isolation a challe ...

Blue native gel electrophoresis (BNGE) is a powerful tool for analyzing native protein complexes from biological membranes as well as water-soluble proteins. It can be used for determining relative molecular masses of protein complexes and their subunit composition and for the detec ...

Import of proteins into mitochondria is a pivotal process in the biogenesis of mitochondria. Only about 1% of the 1000–2000 different proteins constituting the mitochondrion are encoded in the mitochondrial deoxyribonucleic acid (DNA). All others are specified by nuclear genes. They a ...

In plants, the majority of mitochondrial and chloroplast proteins are nuclear encoded, synthesized on cytosolic polyribosomes, and then imported into the organelle. Most of the nuclear encoded precursor proteins contain an N-terminal extension called signal or targeting pepti ...