细胞功能测定

Somatic cell nuclear transfer (SCNT) is a technically and biologically challenging procedure during which a differentiated committed nucleus undergoes rapid reprogramming into the totipotent state in a few hours. SCNT can be utilized to generate patient- and disease-specific em ...

Deepening our knowledge on the regulation of the plant cell division cycle depends on techniques that allow for the enrichment of cell populations in defined cell cycle phases. Synchronization of cell division can be achieved using different plant tissues; however, well-established c ...

When removed from the follicles, during the 44h process of in vitro maturation (IVM) fully grown porcine oocytes resume meiosis spontaneously from the late diplotene stage of the first meiotic prophase and proceed to the metaphase-II (MII) stage at which they remain arrested until fertiliz ...

Protozoans are single-cell eukaryotes and many of the best studied protozoans are parasitic to humans (e.g., Plasmodium falciparum causing malaria and Trypanosoma brucei causing sleeping sickness). These organisms are distantly related to humans but with retained eukaryotic ty ...

The budding yeast Saccharomyces cerevisiae and fission yeast Schizosaccharomyces pombe are amongst the simplest and most powerful model systems for studying the genetics of cell cycle control. Because yeast grows very rapidly in simple and economical media, large numbers of cells can ...

This protocol defines conditions under which the germination of spores can be used to synchronize Bacillus subtilis cells, utilizing the time-ordered sequence of events taking place during the transition from spore to vegetative cells. The transition stages involve: phase change, s ...

HeLa is one of the oldest and most commonly used cell lines in biomedical research. Owing to the ease of which they can be effectively synchronized by various methods, HeLa cells have been used extensively for studies of the cell cycle. Here we describe several protocols for synchronization of HeLa c ...

During malignant transformation, cells can increase their ploidy and hence become polyploid (mostly tetraploid). Frequently, however, tetraploid cells undergo asymmetric divisions, in turn entailing a reduction in ploidy and the acquisition of a pseudo-diploid, aneuploid st ...

This chapter will describe the preparation and use of a semipermeabilized (SP) cell system that reconstitutes the initial stages in the assembly and modification of proteins entering the secretory pathway (1). The procedure involves treating cells grown in culture with the detergent d ...

Intermolecular binding forces are intrinsic property of cohesive structures and should be used as the main quantitative criteria for assessing and defining their functional contribution to the maintenance of the anatomical integrity of the adult and embryonal multicellular or ...

The examination of ECM macromolecules by conventional transmission electron microscopy (TEM) or scanning transmission EM (STEM) can provide important information on macromolecular organization and interactions.

The primary functional role of collagen is as a supporting tissue and it is now well established that the aggregated forms of the collagen monomers are stabilized to provide mechanical strength by a series of intermolecular crosslinks. These links are formed by oxidative deamination of the ε- ...

Recent developments in the application of eukaryotic recombinant protein techniques have provided new tools with which to dissect and map functional activities in basement membrane glycoproteins. This has been particularly valuable in the case of laminins where the relationsh ...

For recombinant expression of extracellular matrix (ECM) proteins or their individual domains, the use of transformed mammalian cells offers two major advantages. First, eukaryotic expression can be expected under optimum conditions to produce a large proportion of correctly fo ...

The collagens, proteoglycans, and glycoproteins of the extracellular matrix (ECM) are a diverse collection of macromolecules, heterogeneous in size, structure, and function. However, their primary structures have revealed that their polypeptide components are frequently c ...

Integrins are heterodimeric cell-surface receptors involved in a variety of functions such as binding to the ECM, regulation of the cellular organization, cell migration, proliferation, differentiation, and gene expression (1,2). Integrins in their native environment can diff ...

Collagen fibrils in tissue are generally heterotypic with more than one type of collagen molecule incorporated into the fibril structure. Furthermore specific macromolecules are bound onto the fibril surface influencing both the assembly and the interaction of the fibril with the s ...

Extracellular matrices (ECM) contain a mixture of fibrillar and nonfibrillar macromolecular components, which together form a composite structure (1–3). It is the ECM that defines the architecture, the form, and the biomechanical properties of different tissues (4,5). Among the nonf ...

Genetic disorders of cartilage (chondrodysplasias) are a clinically and genetically heterogeneous group of diseases ranging in severity from relatively mild to severe and lethal forms (1-2). There are over 100 unique well-characterized chondrodysplasia phenotypes and rema ...

The analysis of phenotypes caused by null and mutant alleles is a very powerful means to understand gene function in vivo. Historically, this experimental approach has been widely and successfully used in invertebrate models. Now, thanks to the gene-targeting technology in ES cells, the gen ...