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细胞功能测定

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Differentiation of Human Embryonic Stem Cells Toward the Chondrogenic Lineage

Human embryonic stem cells (hESCs) have the ability to self-replicate and differentiate into cells from all three embryonic germ layers, thereby holding great promise for tissue regeneration applications. However, controlling the differentiation of hESCs and obtaining homog ...

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Feeder-Layer Free Culture System for Human Embryonic Stem Cells

Human embryonic stem cells (hESCs) are pluripotent stem cells derived from the inner cell mass of the blastocyst. Due to their unique properties, hESCs might be used for research fields such as self-renewal, specific lineage differentiation, human developmental biology, and teratolo ...

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A Controlled-Cooling Protocol for Cryopreservation of Human and Non-Human Primate Embryonic Stem Cells

Freeze storage of human embryonic stem (hES) cells has not proven effective using the methods employed for mouse ES (mES) cells, while rhesus ES (rhES) cells are only modestly effectively frozen using common mES cell methods. Because human and rhES cells are passaged and frozen in clusters that ap ...

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Digital Imaging of Stem Cells by Electron Microscopy

This chapter deals with basic techniques of scanning and transmission electron microscopy applicable to stem cell imaging. It is sometimes desirable to characterize the fine structure of embryonic and adult stem cells to supplement the images obtained by phase-contrast and confocal ...

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Cell Surface Markers in Human Embryonic Stem Cells

The pluripotent nature of human embryonic stem cells (hESCs) is based on their potential to form every cell type in the body. Prior to use in directed differentiation strategies, these cells need to be thoroughly characterized. The large number of glycoproteins and carbohydrates that exist on ...

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Generation of a Monoclonal Antibody Library Against Human Embryonic Stem Cells

Differentiated cell types derived from human embryonic stem cells (hESCs) may serve in the future to treat various human diseases and to model early human embryonic development in vitro. Fulfilling this potential, however, requires extensive development of methods and reagents for st ...

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Analytical Methods for Cancer Stem Cells

The primary characteristics of adult stem cells are maintaining prolonged quiescence, ability to self-renew and plasticity to differentiate into multiple cell types. These properties are evolutionarily conserved from fruit fly to humans. Similar to normal tissue repair in organ ...

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Micro RNA Profiling: An Easy and Rapid Method to Screen and Characterize Stem Cell Populations

MicroRNAs (miRNAs) are small regulatory RNAs varying in length between 20 and 24 nucleotides. They are thought to play a key role during development by negative gene regulation at the post-transcriptional level. Recent studies using quantitative polymerase chain reaction (QPCR) and n ...

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Gene Transfer Via Nucleofection Into Adult and Embryonic Stem Cells

The use of embryonic and adult stem cells as therapeutic agents is gaining momentum. A major impediment in the use of stem cells for genetic disorders is their ability to undergo genetic modification. The recognition of various site-specific integration methods open up a new avenue for gene the ...

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Role of Natural Killer Cells and Killer Immunoglobulin-Like Receptor Polymorphisms: Association of HLA and KIRs

Natural killer cells play an important role in innate immunity. They act against infected and transformed cells as part of the immune surveillance process. Their interactions with the human leukocyte antigens (HLAs) create a situation where they may act against donor hematopoietic cel ...

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Identification of Bone Marrow Derived Nonhematopoietic Cells by Double Labeling with Immunohistochemistry and In Situ Hybridization

Stem cell migration/trafficking is a field of interest that is shared by pathologists, histologists, clinical transplantation teams, cardiologists, neurologists, and many other members of different disciplines. Until the findings of a successful combination of in situ methods, ...

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Molecular Methods Used for Detection of Minimal Residual Disease Following Hematopoietic Stem Cell Transplantation in Myeloid Disorders

Monitoring of minimal residual disease (MRD) in patients with acute or chronic myeloid disorders is routinely performed after allogeneic or autologous transplantation. The detection of MRD helps to identify patients who are at high risk for leukemic relapse after transplantation. ...

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Detection of Impending Graft Rejection and Relapse by Lineage-Specific Chimerism Analysis

Molecular surveillance of hematopoietic chimerism has become part of the routine diagnostic program in patients after allogeneic stem cell transplantation. Chimerism testing permits early prediction and documentation of successful engraftment, and facilitates early d ...

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Molecular Methods Used for The Detection of Autologous Graft Contamination in Lymphoid Disorders

Intensified treatments aimed at maximal tumor reduction are an important therapeutic option for patients affected by B-cell malignancies. The possibility of obtaining a relevant number of clinical complete remissions after these treatments prompted the application of molec ...

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Application of Proteomics to Posttransplantational Follow-Up

Proteomic screening of complex biological samples becomes of increasing importance in clinical research and diagnosis. It is expected that the meager number of approx 35,000 human genes gives rise to more than 1,000,000 functional entities at the protein level. Thus, the proteome provid ...

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Molecular Profiling of Hematopoietic Stem Cells

Gene expression profiling using microarrays is a powerful method for studying the biology of hematopoietic stem cells (HSCs). Here, we present methods for activating HSCs with the chemotherapeutic drug 5-Fluorouracil, isolating HSCs from whole bone marrow, and performing microar ...

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Methods for Imaging Cell Fates in Hematopoiesis

Modern imaging technologies that allow for in vivo monitoring of cells in intact research subjects have opened up broad new areas of investigation. In the field of hematopoiesis and stem cell research, studies of cell trafficking involved in injury repair and hematopoietic engraftment h ...

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An Overview of HLA Typing for Hematopoietic Stem Cell Transplantation

Selection of a related or unrelated haematopoietic stem cell donor for a patient requires accurate matching of human leukocyte antigen (HLA) genes in order to maximise the beneficial effects of the transplant. There are a number of different approaches that can be made in order to achieve HLA type ...

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Sequence-Specific Primed PCR (PCR-SSP) Typing of HLA Class I and Class II Alleles

For donor selection in hematopoetic stem cell transplantation, two-digit sequence-specific oligonucleotide (SSO) typing may be sufficient in the related sibling transplant setting. However, SSO typing is not sensitive enough to differentiate between the alleles that share the ...

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HLA Typing with Sequence-Specific Oligonucleotide Primed PCR (PCR-SSO) and Use of the Luminex Technology

The hybridization products obtained by PCR using sequence-specific oligonucleotides (PCR-SSO) can be traced either by colorimetric-(streptavidin-biotin), X-ray-(digoxigenin-CSPD), or fluorescence-(FITC, PE) based detection systems. To achieve a faster, reliable, aut ...

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