细胞技术

FlyBase (http://flybase.org) is the primary database of integrated genetic and genomic data about the Drosophilidae, of which Drosophila melanogaster is the most extensively studied species. Information in FlyBase originates from a variety of sources ranging from large-scale ge ...

Over the past decade the adoption and refinement of the GAL4 system by the Drosophila field has resulted in a wide array of tools with which the researcher can drive transgene expression in a precise spatiotemporal pattern. The GAL4 system relies on two components: (1) GAL4, a transcriptional acti ...

The development of a technique to stably integrate exogenous DNA into the germline of Drosophila melanogaster marked a milestone in the ability to study gene function in the fly. On the molecular level germline transformation mainly relies on a particular transposable element, the D. mela ...

Mobile elements were first used as a mutagenesis tool that introduces a molecular tag in the genes of interest. This facilitated subsequent molecular cloning and eventually promoted molecular analysis of a large number of fly genes. Soon after, P-elements were modified to detect genes not on ...

The success of Drosophila as a genetic model organism is based on the efficient generation, recovery, and identification of new mutations. Various agents have been used to induce de novo DNA lesions. However, the use of mutagenic alkylating agents, especially ethyl methanesulfonate (EMS), ...

RNA interference (RNAi) has become a widely used tool to analyze biological functions in vivo and in vitro. With the availability of an increasing number of Drosophila cell lines, a variety of different processes can be studied ranging from cell cycle control defects to signaling pathway acti ...

We present detailed protocols for two methods of gene targeting in Drosophila. The first, ends-out targeting, is identical in concept to gene replacement techniques used routinely in mammalian and yeast cells. In Drosophila, the targeted gene is replaced by the marker gene white + (although o ...

There is a need for direct imaging of effects on tumour vasculature in assessment of response to anti-angiogenic drugs and vascular disrupting agents. Imaging tumour vasculature depends on differences in permeability of vasculature of tumour and normal tissue, which cause changes in p ...

Tumour neovascularization acquires vessels through a number of processes, including angiogenesis, vasculogenesis, vascular remodelling, intussusception, and possibly vascular mimicry in certain tumours. The end result of the tumour vasculature has been quantified by co ...

Abstract The field of lymphatic research has benefited enormously from the recent discovery of “marker” proteins that permit not only the identification and quantitation of lymphatic vessels in tissue sections for tumour pathology but also the isolation of primary lymphatic endot ...

The endothelium residing in different vascular beds displays high-degree phenotypic heterogeneity at morphological, functional, biochemical, and molecular levels. Endothelial cells (ECs) can be easily harvested from large vessels by mechanical removal or collagenase di ...

Smooth muscle cells (SMCs) are key players in the pathogenesis of atherosclerosis and restenosis; however, they are also important in formation and development of de novo blood vessels during vasculogenesis and angiogenesis. Vascular SMCs can be formed by proliferation of existing SM ...

This chapter describes detailed methods for the isolation of primary human lymphatic endothelial cells from neonatal foreskin. We also provide protocols and information for their characterization and propagation. Isolation of primary human lymphatic endothelial cells req ...

Recent studies have found that bone marrow-derived cells give rise to endothelial cells during states of tissue repair and disease. We have found that one key integrin, integrin-α4β1, promotes the homing of circulating endothelial progenitor cells (EPCs) to sites of ongoing tissue repai ...

Neovascularization plays a role in several pathological conditions, including tumor growth, arthritis, and choroidal neovascularization. Investigators from different fields can choose from several available angiogenesis assays according to their specific needs. Th ...

Angiogenesis is a complex sequential process involving endothelial activation, basement membrane degradation, endothelial sprouting from the parent vessel, invasion of the extracellular matrix, endothelial proliferation, vessel elongation, branching, anastomo ...

Methods are described for analysing adhesion of isolated cells (such as leucocytes, tumour cells, or precursor cells) to purified adhesion receptors or cultured endothelial cells. “Static” assays (in which cells are allowed to settle on the adhesive substrates) and flow-based assays ( ...

The corneal endothelium plays a key role in the physiology of the cornea, maintaining its transparency by regulating corneal hydration. Moreover, corneal endothelial cells play the central role in irreversible corneal graft rejection as human corneal endothelial cells are predom ...

In our efforts aimed at studying the cellular responses to injury, including the angiogenesis of wound healing, we have developed a novel three-dimensional (3D) skin equivalent that is comprised of multiple cell types found in normal human skin or chronic wound beds. The in vitro model contains a ...

Successful therapeutic angiogenesis requires an understanding of how the milieu of growth factors available combine to form a mature vascular bed. This requires a model in which multiple physiological and cell biological parameters can be identified. The adenoviral-mediated me ...