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细胞功能测定

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Techniques in Embryoid Body Formation from Human Pluripotent Stem Cells

Embryoid bodies (EBs) can be generated by culturing human pluripotent stem cells in ultra-low attachment culture vessels, under conditions that are adverse to pluripotency and proliferation. EBs generated in suspension cultures are capable of differentiating into cells of the ect ...

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Formation of Embryoid Bodies from Human Pluripotent Stem Cells Using AggreWell Plates

Many human embryonic stem (hES) and induced pluripotent stem (hiPS) cell differentiation protocols begin with the formation of three-dimensional aggregates of cells called embryoid bodies (EBs). Traditional EB formation methods result in a heterogeneous population of EB sizes a ...

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Feeder-Independent Culture Systems for Human Pluripotent Stem Cells

The continued success of pluripotent stem cell research is ultimately dependent on access to reliable and defined reagents for the consistent culture and cryopreservation of undifferentiated, pluripotent cells. The development of defined and feeder-independent culture me ...

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Methods to Culture, Differentiate, and Characterize Neural Stem Cells from the Adult and Embryonic Mouse Central Nervous System

Since the discovery of neural stem cells (NSC) in the embryonic and adult mammalian central nervous system (CNS), there have been a growing numbers of tissue culture media and protocols to study and functionally characterize NSCs and its progeny in vitro. One of these culture systems introduced ...

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Eradication of Mycoplasma Contaminations

Mycoplasma contaminations have a multitude of effects on the cultured cell lines that may influence the results of experiments or pollute bioactive substances used in human medicine. The elimination of mycoplasma contaminations of cell cultures has become a practical alternative ...

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Hepatic Differentiation of Embryonic Stem Cells by Murine Fetal Liver Mesenchymal Cells

Hepatocytes derived from embryonic stem cells (ESCs) are a potential cell source for regenerative medicine. However, it has been technically difficult to differentiate ESCs into mature hepatocytes because the definitive growth factors and molecular mechanisms governing hep ...

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Isolation and Culture of Skeletal Muscle Myofibers as a Means to Analyze Satellite Cells

Multinucleated myofibers are the functional contractile units of skeletal muscle. In adult muscle, mononuclear satellite cells, located between the basal lamina and the plasmalemma of the myofiber, are the primary myogenic stem cells. This chapter describes protocols for isolat ...

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Cocultivation of Human Oral Keratinocytes and Human Osteoblast-Like Cells

Head and neck reconstruction transplants often require a bony structure but also tissue for the intraoral lining. This is why oral keratinocytes and osteoblast-like cells are essential cell types for combined tissue engineered transplants for defects in the field of craniomaxillof ...

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Isolation and Characterization of Human Hair Follicle Epithelial Cells

The mammalian hair follicle epithelial component contains various lineages of keratinocytes as well as their progenitor/stem cells. To characterize the subpopulations contained within this component and assess their functional capacity, the development of a feasible meth ...

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Enzymatic Dissociation, Flow Cytometric Analysis, and Culture of Normal Mouse Mammary Tissue

Evidence is emerging that the mouse mammary epithelium is arranged as a hierarchy that spans from stem cells to lineage-restricted progenitor cells to differentiated luminal and myoepithelial cells. The use of fluorescence-activated cell sorting (FACS) in combination with quant ...

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Human Prostate Epithelial Cell Cultures

Prostate cancer is the most common male cancer in the United States. Research on the mechanisms of prostate cancer progression has been limited by the lack of suitable in vitro systems. A hurdle in understanding the molecular genetic changes in prostate cancer has been the difficulty in establi ...

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In Vitro Methods to Culture Primary Human Breast Epithelial Cells

Current evidence suggests that much like leukemia, breast tumors are maintained by a small subpopulation of tumor cells that have stem cell properties. These cancer stem cells are envisaged to be responsible for tumor formation and relapse. Therefore, knowledge about their nature will pr ...

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Generation of a Pool of Human Platelet Lysate and Efficient Use in Cell Culture

Human platelets represent a promising source of bioactive substances as growth factors not just for in vivo wound healing and tissue repair, but also for the expansion of human stem and progenitor cells in vitro. The replacement of fetal bovine serum (FBS) as a standard culture supplement by human ...

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Isolation and Culture of Mesenchymal Stem Cells from Mouse Compact Bone

The bone marrow (BM) of numerous species, including rodents and man, contains a rare population of cells termed marrow stromal cells or mesenchymal stem cells (MSC). Given the ability of these cells to differentiate into cells of the osteogenic, chondrogenic and adipogenic lineages, there is ...

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Isolation, Enumeration, and Expansion of Human Mesenchymal Stem Cells in Culture

Human bone marrow (BM) contains a population of non-hematopoietic stem cells also termed stromal cells, mesenchymal cells or multipotent mesenchymal stromal cells (MSCs). These cells have unique stem cell-like properties including their ability to self-renew, differentiate in ...

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Detection of Mycoplasma Contaminations

Mycoplasma contamination of cell lines is one of the major problems in cell culture technology. The �specific, sensitive, and reliable detection of mycoplasma contamination is an important part of mycoplasma control and should be an established method in every cell culture laboratory. ...

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Generation and Establishment of Murine Adherent Cell Lines

We describe a method to derive cell lines and clones from cells of the murine midgestation aorta-gonads-mesonephros (AGM) microenvironment. We start from subdissected AGM regions in “explant” or “single cell suspension” type cultures from embryos transgenic for tsA58, a temperature ...

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Studying Leukocyte Recruitment Under Flow Conditions

Leukocyte recruitment from the vasculature occurs under conditions of haemodynamic shear stress. The parallel plate flow chamber apparatus is an in vitro system that is widely used to study leukocyte recruitment under shear conditions. The flow chamber is a versatile tool for examining ...

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Colony Forming Cell Assays for Human Hematopoietic Progenitor Cells

Hematopoietic stem cells (HSCs) present in small numbers in adult bone marrow (BM), peripheral blood (PB) and umbilical cord blood (CB) produce a heterogeneous pool of progenitors that can be detected in vitro using colony forming cell (CFC) assays. Hematopoietic progenitor cells prolif ...

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Long-Term Culture-Initiating Cell Assay for Mouse Cells

The long-term culture-initiating cell (LTC-IC) assay is a well-established in vitro assay used to enumerate primitive mouse hematopoietic stem cells (HSCs) and relies on the two cardinal functions of HSCs: ability to self-renew and differentiation capacity. LTC-ICs present in minima ...

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