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细胞功能测定

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Transplantation of Oligodendrocyte Progenitor Cells in Animal Models of Leukodystrophies

Leukodystrophies represent a wide variety of hereditary disorders of the white matter in the central nervous system, where the patients, mostly in infancy or childhood, suffer from progressive and often fatal neurological symptoms due to either a delay or lack of myelin development or loss ...

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Animal Models of Multiple Sclerosis

Since its first description, experimental autoimmune encephalomyelitis, originally designated experimental allergic encephalitis (EAE), has been proposed as animal model to investigate pathogenetic hypotheses and test new treatments in the field of central nervous sys ...

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Animal Models of Neurodegenerative Diseases

Animal models of neurodegenerative disease are excellent tools for studying pathogenesis and therapies including cellular transplantation. In this chapter, we describe different models of Huntington’s disease and Parkinson’s disease, stereotactic surgery (used in crea ...

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Human Trials for Neurodegenerative Disease

The lack of disease-modifying treatments currently available for not just some but most neurodegenerative diseases, including Parkinson’s disease, Huntington’s disease, and even stroke, helps explain increasing interest in cell-based therapies. One key aim of such treatment is ...

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Adult Stem Cells for the Treatment of Neurological Disease

The characteristic CNS responses to injury including increased cell production and attempts at regenerative repair – implicitly predicted where not directly demonstrated by Cajal, but only now more fully confirmed – have important implications for regenerative therapies. Spo ...

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Neural Transplantation and Stem Cells

Recent results have raised important questions on our ability to amplify stem cell populations in sufficient numbers as to be useful for therapy. Several reports have indicated that human stem cell populations harvested from the adult have low or undetectable telomerase levels, age in cul ...

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A Rat Middle Cerebral Artery Occlusion Model and Intravenous Cellular Delivery

A useful experimental model to study the pathophysiology of cerebral ischemia without craniectomy is the middle cerebral artery occlusion (MCAO) model. In this model, an intraluminal suture is advanced from the internal carotid artery to occlude the base of the MCA. Standardized proced ...

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MR Tracking of Stem Cells in Living Recipients

Noninvasive cellular imaging allows the real-time tracking of grafted cells as well as the monitoring of their migration. Several techniques for in vivo cellular imaging are available that permit the characterization of transplanted cells in a living organism, including magnetic r ...

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Identifying Neural Progenitor Cells in the Adult Brain

There is incontrovertible evidence that neural progenitor cells (NPC) are found in the adult brain. The ability to identify and track NPC in the adult brain is of considerable importance if the properties of these cells are to be harnessed as potential therapies for degenerative brain disorde ...

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Immune Ablation Followed by Autologous Hematopoietic Stem Cell Transplantation for the Treatment of Poor Prognosis Multiple Sclerosis

Complete abrogation of the inflammatory response by high-dose cytotoxic therapy at an early stage of MS, when the nervous system has not yet sustained irreparable damage may be successful at preventing the inexorable progression. Immunological and hematological reconstitution ...

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Differentiation of Neuroepithelia from Human Embryonic Stem Cells

We describe the method for efficiently differentiating human embryonic stem cells to neuroepithelial cells in a chemically defined condition. The protocol was established based on the fundamental principle of in vivo neuroectodermal development. The temporal course, morpho ...

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Derivation of High-Purity Oligodendroglial Progenitors

Oligodendrocytes are a type of glial cells that play a critical role in supporting the central nervous system (CNS), in particular insulating axons within the CNS by wrapping them with a myelin sheath, thereby enabling saltatory conduction. They are lost, and myelin damaged – demyelination – in a ...

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Flow Cytometric Characterization of Neural Precursor Cells and Their Progeny

It is now clear that the adult central nervous system contains a population of neural stem and progenitor cells which act as a reservoir to underpin cell genesis for the lifetime of the animal. Unfortunately, understanding how these cells are activated both under normal conditions and followi ...

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Evaluation of Rapamycin-Induced Cell Death

Mammalian target of rapamycin (mTOR) is an evolutionarily conserved kinase that integrates signals from nutrients and growth factors for the coordinate regulation of many cellular processes, including proliferation and cell death. Constitutive mTOR signaling characteri ...

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Detection of Cytoplasmic and Nuclear Functions of mTOR by Fractionation

Subcellular localization constitutes the environment in which proteins act. It tightly controls access to and availability of different types of molecular interacting partners and is therefore a major determinant of protein function and regulation. Originally thought to be a mere ...

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Overexpression or Downregulation of mTOR in Mammalian Cells

This chapter presents an overview of the methods that have been used to overexpress or downregulate the level of mTOR isoforms in mammalian cells. The techniques of transient overexpression, generation of stable cell lines, retroviral- and lentiviral-mediated overexpression or do ...

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An In Vitro Assay for the Kinase Activity of mTOR Complex 2

The mTOR complex 2 (mTORC2) is a protein kinase complex involved in many important physiological processes through the regulation of its substrates, such as Akt, SGK1, and conventional PKC. Its activity is modulated negatively by interaction with DEPTOR and positively by the TSC1–TSC2 pro ...

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Isolation of the mTOR Complexes by Affinity Purification

The mammalian Target Of Rapamycin (mTOR) protein is a central component of the essential and highly conserved signaling pathway that emerged as a critical effector in regulation of cell physiology. Biochemical studies defined mTOR as the protein kinase that exists at least in two distinct c ...

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mTOR Activity Under Hypoxia

The adaptive response to hypoxia, low oxygen tension, involves inhibition of energy-intensive cellular processes including protein translation. This effect is mediated in part through a decrease in the kinase activity of mammalian target of rapamycin complex 1 (mTORC1), a master reg ...

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Evaluation of the Nutrient-Sensing mTOR Pathway

mTOR, an evolutionarily conserved Ser/Thr protein kinase, belongs to the PI3K-related kinase family, which also includes DNA-PKcs, ATM, and ATR. Although other PI3K-related kinase family members have been shown to secure genomic integrity by sensing DNA damage and related stresses, mT ...

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