细胞功能测定

Microinjection is an excellent technique for studying the acute responses of cells to proteins and can be used to investigate the effects of mutations in proteins on their activity. It has been used widely to study the responses to Rho family guanosine triphosphatases and is particularly use ...

Some membrane proteins must be clustered at target sites to efficiently perform their functions. PDZ domain-containing scaffold proteins bind to the tails of target membrane proteins and promote their localization and clustering on the cell surface. This chapter describes the expe ...

The recognition that Rho guanosine triphosphatases (GTPases) (Rho, Rac, and Cdc42) play important regulatory roles in many areas of cell biology has made the ability to measure their activity in cells an important biological tool. Because Rho GTPases become activated by conversion from g ...

Phospholipase D (PLD) enzymes are present in all animal and plant species and have been linked to many critical cellular processes, including proliferation, differentiation, motility, and secretion. The functional significance of PLD derives from its generation of phosphatidic a ...

Phospholipase D (PLD) hydrolyzes phosphatidylcholine to produce the membraneassociated second messenger, phosphatidic acid (PA) and choline. Two phospholipase D enzymes—PLD1 and PLD2—have been identified, although their regulatory mechanisms are yet to be fully underst ...

Microarrays are made by immobilizing to a solid support thousands of DNA probes that detect soluble complementary target sequences using the hybridization pairing rules of nucleic acids. Receptor triggering induces a cascade of signaling events that often involves the modulation ...

Introducing genes into cells by retroviral transduction has greatly increased the abil-ity to study signal transduction pathways in primary cells. Retroviral transduction has proven to be an efficient method to express genes of interest in cells that are difficult to manipulate using ...

Proteomics is the study of the set of proteins, or proteome, expressed by a cell under specific conditions. Proteomics methodology consists of protein extraction, protein separation, and protein identification. Currently, two-dimensional gel electrophoresis (2DE) and matrix- ...

Stromal-derived factor (SDF)-1, an α-chemokine that binds to G protein-coupled seven transmembrane-spanning receptor, CXCR4, plays an important and unique role in regulating the trafficking of normal hematopoietic stem/progenitor cells and their homing/ retention in bone mar ...

CXCR4, the receptor for stromal cell-derived factor (SDF)-1, was expressed in Saccharomyces cerevisiae coupled to the pheromone response pathway via a chimeric Gα subunit. Engagement of CXCR4 by SDF-1 resulted in expression of reporter genes, HIS3 or lacZ under the transcriptional cont ...

The ability of G protein-coupled receptors to transduce signaling typically is induced by the binding of an appropriate ligand (agonist), resulting in a conformational change of the receptor and the subsequent interaction with the G protein heterotrimer. Some mutants of G protein-cou ...

G protein-coupled receptors are involved in the regulation of many aspects of normal physiology and pathology. Recent research has broadened our view of how the cell trans-duces ligand binding to cellular responses. It is becoming clear that phenomena that take place both at the cell surface, ...

Seven transmembrane-spanning receptors, widely referred to as G protein-coupled receptors (GPCRs), mediate a broad spectrum of extracellular signals at the plasma membrane through G proteins, thereby modulating a variety of biological processes. In addition to G proteins, they al ...

Traditionally, lipid rafts have been defined by their insolubility in ice-cold Triton X-100 and low-buoyant density. These low-density membrane microdomains have been referred to as detergent-resistant membranes, Triton-insoluble membranes, and Tritoninsoluble floati ...

We have developed protocols whereby a total of 30–90 � 106 hMSCs with an average viability greater than 90% can be produced in a single multilevel Cell Factory from a relatively small (1–3 mL) bone marrow aspirate in 14–20 d. It is possible to generate as many as 5 � 108 multipotent stromal cells (MSCs) from a single ...

Mesenchymal stem cells (MSCs), sometimes referred to as marrow stromal cells or multipotential stromal cells, represent a class of adult progenitor cells capable of differentiation to several mesenchymal lineages. They can be isolated from many tissues although bone marrow has been u ...

The STRO-1 antibody can be used as a single reagent to isolate human bone marrow stromal stem cells (BMSSC), owing to its restricted specificity to a cell surface molecule expressed by clonogenic BMSSC, with little or no reactivity to hematopoietic stem/progenitor populations or mature str ...

Human adipose tissue has been shown to contain a population of cells that possesses extensive proliferative capacity and the ability to differentiate into multiple cell lineages. These cells are referred to as adipose tissue-derived stem cells (ADSCs) and are generally similar, though ...

The rapid development of microarray technology during the last decade has greatly expanded the ability to define the genes expressed in cells. This chapter will focus on describing the steps required for conducting successful microarray experiments with multipotential stromal c ...

Adipose tissue has proven to serve as an abundant, accessible, and rich source of adult stem cells with multipotent properties suitable for tissue engineering and regenerative medical applications. Here, we describe a detailed method for the isolation and expansion of adipose-deriv ...