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细胞功能测定

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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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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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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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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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Monitoring Spindle Assembly and Disassembly in Yeast by Indirect Immunofluorescence

The budding yeast Saccharomyces cerevisiae has become a laboratory favorite for the study of several microtubule-dependent processes, including assembly of the mitotic spindle and spindle positioning. This is owing not only to the pliancy of its genetic system but also to the simplici ...

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Assay of Deoxyhypusine Hydroxylase Activity

The eukaryotic translation initiation factor 5A (eIF5A) is the only cellular protein that contains an unusual amino acid, hypusine . eIF5A and its hypusine/deoxyhypusine modification are vital for eukaryotic cell proliferation. Hypusine is formed posttranslationally by two en ...

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Identification and Assay of Allosteric Regulators of S-Adenosylmethionine Decarboxylase

Polyamine biosynthesis is extensively regulated in cells by multiple mechanisms, including regulation of enzyme activity posttranslationally. The identified regulatory factors include both small molecules and regulatory proteins, and the mechanisms vary in different ...

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Protocols for Studying Antizyme Expression and Function

Antizyme (AZ) is a key molecule in feedback regulation of cellular polyamines. It is induced by polyamines through stimulation of ribosomal frameshifting during its translation. In mammals, AZ is diverged into three paralogs, AZ1–3. Tissue and subcellular distribution are different ...

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Identification, Assay, and Functional Analysis of the Antizyme Inhibitor Family

Polyamines are small aliphatic polycations present in all living cells. Polyamines are involved in regulating fundamental cellular functions and are absolutely essential for the process of cellular proliferation. Because they fulfill essential cellular functions, their i ...

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Posttranscriptional Regulation of Ornithine Decarboxylase

Activity of the polyamine biosynthetic enzyme ornithine decarboxylase (ODC) and intracellular levels of ODC protein are controlled very tightly. Numerous studies have described ODC regulation at the levels of transcription, translation, and protein degradation in normal cel ...

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Identification and Assays of Polyamine Transport Systems in Escherichia coli and Saccharomyces cerevisiae

Polyamine content in cells is regulated by biosynthesis, degradation, and transport. With regard to transport, uptake and excretion proteins exist in Escherichia coli and Saccharomyces cerevisiae. In E. coli, the uptake systems comprise a spermidine-preferential uptake system c ...

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Heparan Sulfate Proteoglycan-Mediated Polyamine Uptake

The polyamines are polycationic compounds essential for cellular proliferation and transformation. In addition to a well-defined biosynthesis pathway, polyamines are internalized into cells by as yet incompletely defined mechanisms. Numerous reports have shown that effi ...

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