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Analysis of Meiotic Recombination in Caenorhabditis elegans

Caenorhabditis elegans is an important experimental organism for the study of recombination during meiosis. A variety of techniques have been developed for the measurement of meiotic recombination in C. elegans, ranging from traditional genetic measures to direct cytological d ...

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Genetic Analysis of Meiotic Recombination in Schizosaccharomyces pombe

The fission yeast Schizosaccharomyces pombe is well-suited for studying meiotic recombination. Methods are described here for culturing S. pombe and for genetic assays of intragenic recombination (gene conversion), intergenic recombination (crossing-over), and spore via ...

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Measurement of Spatial Proximity and Accessibility of Chromosomal Loci in Saccharomyces cerevisiae Using Cre/loxP Site-Specific Recombination

Several methods have been developed to measure interactions between homologous chromosomes during meiosis in budding yeast. These include cytological analysis of fixed, spread nuclei using fluorescence in situ hybridization (FISH) (1, 2), visualization of GFP-labeled chrom ...

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Modulating and Targeting Meiotic Double-Strand Breaks in Saccharomyces cerevisiae

Meiotic recombination is initiated by DNA double-strand breaks (DSBs) formed by the evolutionary conserved Spo11 protein. Along the S. cerevisiae chromosomes, the DSB sites are not evenly distributed and the cleavage frequencies vary 10–100-fold from site to site. Herein are reviewed t ...

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Methods for Analysis of Crossover Interference in Saccharomyces cerevisiae

Interest in crossover interference in yeast has been spurred by the discovery and characterization of mutants that alter it as well as by the development and testing of models to explain it. This chapter describes methods for detecting and for measuring interference, with emphasis on those th ...

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Optimizing Sporulation Conditions for Different Saccharomyces cerevisiae Strain Backgrounds

Different sporulation and pre-sporulation regimens were compared for a number of commonly used laboratory strains of S. cerevisiae to define conditions that support high-efficiency sporulation.

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Interaction of Genetic and Environmental Factors in Saccharomyces cerevisiae Meiosis: The Devil is in the Details

One of the most important principles of scientific endeavour is that the results be reproducible from lab to lab. Although research groups rarely redo the published experiments of their colleagues, research plans almost always rely on the work of someone else. The assumption is that if the same e ...

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Regeneration of Skin and Cornea by Tissue Engineering

Progress in tissue engineering has led to the development of technologies allowing the reconstruction of autologous tissues from the patient’s own cells. Thus, tissue-engineered epithelial substitutes produced from cultured skin epithelial cells undergo long-term regene ...

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Bone Marrow-Derived Mesenchymal Stem Cells: Isolation, Expansion, Characterization, Viral Transduction, and Production of Conditioned Medium

Mesenchymal stem cells (MSCs) are defined as self-renewing and multipotent cells capable of differentiating into multiple cell types, including osteocytes, chondrocytes, adipocytes, hepatocytes, myocytes, neurons, and cardiomyocytes. MSCs were originally isolated from ...

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Isolation, Propagation, and Characterization of Human Umbilical Cord Perivascular Cells (HUCPVCs)

Current sources of mesenchymal cells, including bone marrow, fat and muscle, all require invasive procurement procedures, and provide relatively low frequencies of progenitors. Here, we describe the non-invasive isolation, and characterization, of a rich source of mesenchymal p ...

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Prospective Isolation of Mesenchymal Stem Cells from Mouse Compact Bone

Bone marrow from numerous species, including rodents and man, has been shown to contain a rare population of cells known as marrow stromal cells or mesenchymal stem cells (MSC). Given the innate ability of these cells to give rise to multiple tissue types including bone, fat and cartilage, there is con ...

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Isolation and Grafting of Single Muscle Fibres

Satellite cells are mononucleate muscle precursor cells resident beneath the basal lamina, which surrounds each skeletal muscle fibre. Normally quiescent in adult muscle, in response to muscle damage satellite cells are activated and proliferate to generate a pool of muscle precur ...

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Differentiation and Dynamic Analysis of Primitive Vessels from Embryonic Stem Cells

Embryonic stem (ES) cells, which are derived from developing mouse blastocysts, have the ability to differentiate into various cell types in vitro. When placed in basal medium with added serum, mouse ES cells undergo a programed differentiation favoring formation of cell types that are fou ...

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Derivation of Contractile Smooth Muscle Cells from Embryonic Stem Cells

Smooth muscle cells (SMCs) play a key role in vascular physiology and pathology. An appreciation of normal SMCs developmental mechanisms will likely lead to a better understanding of disease processes and potentially to novel treatment strategies. We present a method for generating re ...

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Islet-Derived Progenitors as a Source of In Vitro Islet Regeneration

Current therapies do not prevent the complications of diabetes. Furthermore, these therapies do not address the underlying pathology; the lack of functional β-cell mass that occurs in both types 1 and 2 diabetes. While pancreas and islet transplantation do serve to increase β-cell mass, a lack ...

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Isolation and Characterization of Hepatic Stem Cells, or Oval Cells, from Rat Livers

The pace of research on the potential therapeutic uses of liver stem cells or “oval cells” has accelerated significantly in recent years. Concurrent advancements in techniques for the isolation and characterization of these cells have helped fuel this research. Several models now exist ...

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Isolation and Culture of Epithelial Stem Cells

In the skin, epithelial stem cells in the hair follicle contribute not only to the generation of a new hair follicle with each hair cycle, but also to the repair of the epidermis during wound healing. When these stem cells are isolated and expanded in culture, they can give rise to hair follicles, sebaceous g ...

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Transplantation of Embryonic Stem Cell-Derived Dopaminergic Neurons in MPTP-Treated Monkeys

One of the target diseases of cell-replacement therapy is Parkinson’s disease. Clinical experiences with fetal dopaminergic cell graft have shown that the therapy is effective, but limited and accompanied by side effects, such as dyskinesia. So, the therapy needs to be further improved and ...

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Isolation and Manipulation of Mammalian Neural Stem Cells In Vitro

Neural stem cells are potentially a source of cells not only for replacement therapy but also as drug vectors, bringing bioactive molecules into the brain. Stem cell-like cells can be isolated readily from the human brain, thus, it is important to find culture systems that enable expansion in a mult ...

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Detecting the Influence of Cell Cycle Regulatory Proteins on Human Telomeres

Maintenance of genome stability depends on an appropriate response to deoxyribonucleic acid (DNA) damage. This response is based on a complex network of signaling pathways that activate numerous processes and ultimately lead to damage repair and cellular survival or cell death. Thus, a r ...

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