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细胞功能测定

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Cloning Mice and ES Cells by Nuclear Transfer from Somatic Stem Cells and Fully Differentiated Cells

Cloning animals by nuclear transfer (NT) has been successful in several mammalian species. In addition to cloning live animals (reproductive cloning), this technique has also been used in several species to establish cloned embryonic stem (ntES) cell lines from somatic cells. It is the latt ...

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Laboratory Guidelines for Animal Care

Animal research is a controversial subject because of the ethical and moral implications of using unwilling research subjects in potentially painful or distressful procedures usually ending in euthanasia. As such, it must be conducted in a compassionate and responsible manner gear ...

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Manipulating and Imaging the Early Xenopus laevis Embryo

Over the past half century, the Xenopus laevis embryo has become a popular model system for studying vertebrate early development at molecular, cellular, and multicellular levels. The year-round availability of easily fertilized eggs, the embryo’s large size and rapid development, and ...

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Manipulation of Gene Function in Xenopus laevis

Xenopus laevis embryos are particularly well suited to address questions requiring either knockdown or overexpression of genes in a tissue-specific fashion during vertebrate embryonic development. These manipulations are achieved by targeted injection of either antisen ...

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Developmental Genetics in Xenopus tropicalis

The diploid pipid frog Xenopus tropicalis has recently emerged as a powerful new model system for combining genetic and genomic analysis of tetrapod development with embryological and biochemical assays. Its early development closely resembles that of its well-understood tetra ...

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Embryological and Genetic Manipulation of Chick Development

The ability to combine embryological manipulations with gene function analysis makes the chick a valuable system for the vertebrate developmental biologist. We describe methods for those unfamiliar with the chick wishing to initiate chick experiments in their lab. After outlining ...

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Embryological Manipulations in Zebrafish

Due to the powerful combination of genetic and embryological techniques, the teleost fish Danio rerio has emerged in the last decade as an important model organism for the study of embryonic development. It is relatively easy to inject material such as mRNA or synthetic oligonucleotides to re ...

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Essential Techniques for Introducing Medaka to a Zebrafish LaboratoryTowards the Combined Use of Medaka and Zebrafish for Further Genetic Dissection o

The medaka, Oryzias latipes, a small egg-laying freshwater fish, is one of the three vertebrate model organisms in which genome-wide phenotype-driven mutant screens have been carried out. Despite a number of large-scale screens in zebrafish, a substantial number of mutants with new disti ...

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Practical Approaches for Implementing Forward Genetic Strategies in Zebrafish

The tropical fresh water minnow, Danio rerio, more commonly known as zebrafish, has emerged rapidly over the last decade as a powerful tool for developmental geneticists. External fertilization, high fecundity, a short generation time, and optical transparency of embryos during early ...

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Ex Utero Culture and Live Imaging of Mouse Embryos

Mouse genetic approaches when combined with live imaging tools have the potential to revolutionize our current understanding of mammalian biology. The availability and improvement of a wide variety of fluorescent proteins have provided indispensable tools to visualize cells in ...

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Differentiation of Circulating Monocytes into Fibroblast-Like Cells

Monocytes are produced in the bone marrow and enter the blood. They generally leave the blood and enter a tissue, and then become macrophages. In healing wounds, circulating monocytes also enter the tissue and instead of becoming macrophages, can differentiate into fibroblast-like cells c ...

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Generation of a Vascular Niche for Studying Stem Cell Homeostasis

Emerging evidence indicates that endothelial cells (ECs) not only form the passive building blocks of blood vessels that deliver oxygen and nutrients, but also instructively participate in organ regeneration and tumorigenesis by producing tissue-specific angiocrine facto ...

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Enumeration of Very Small Embryonic-Like Stem Cells in Peripheral Blood

In addition to hematopoietic stem and progenitor cells (HSPCs) also other types of stem cells (e.g., �mesenchymal stem cells , endothelial progenitor cells , and very small embryonic-like stem cells ) circulate under steady-state conditions at detectable levels in peripheral blood (PB), ...

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Tracking Inflammation-Induced Mobilization of Mesenchymal Stem Cells

The act of migration is similar for many cell types. The migratory mechanism of mesenchymal stem cells (MSC) is not completely elucidated, yet many of the initial studies have been based on current understanding of the leukocyte migration. A normal function of MSC is the ability of the cell to migrate to ...

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Evaluation of Circulating Endothelial Precursor Cells in Cancer Patients

Results obtained from preclinical studies have shown that endothelial progenitor cells (EPCs) play a crucial role in tumor growth and metastasis. In the clinic, EPCs are present in the peripheral blood of cancer patients in higher numbers than in healthy subjects. These cells are mobilized f ...

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Toxicities of Mobilized Stem Cell Infusion

Infusion of bone marrow or cytokine mobilized stem cells can be associated with severe and potentially life-threatening toxicities. The etiology of the toxicities can be related to volume of infusion, dimethylsulfoxide (DMSO) or bacterial or viral infection of the infused cell produc ...

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Mobilization of Hematopoietic Stem Cells by Depleting Bone Marrow Macrophages

An important factor contributing to hematopoietic stem cell (HSC) mobilization is the ability of mobilizing cytokines and chemotherapy to disturb the cellular components of HSC niches, particularly osteoblasts and their progenitors, and to inhibit the production of HSC support ...

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Combinatorial Stem Cell Mobilization in Animal Models

It has long been recognized that single therapies, such as G-CSF, have a limited capacity to mobilize hematopoietic progenitor cells from the bone marrow. As a consequence in ∼20% of patients insufficient numbers of HPCs are mobilized to perform a bone marrow transplant. Recent studies have sho ...

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Vascular Progenitor Cell Mobilization

Blood vessel formation plays a key role in both physiologic and pathologic tissue growth and healing. Thus, a thorough understanding of the mechanisms underlying neovascularization will translate into innovative clinical treatment strategies for a wide variety of disease proce ...

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Hematopoietic Progenitor Cell Apheresis Processing

Modern apheresis generally results in a mononuclear cell product that is highly enriched for hematopoietic stem and progenitor cells when performed on patients who have mobilized with myeloid growth factors, CXCR4 antagonists or upon recovery from cytotoxic chemotherapy. The dur ...

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