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细胞功能测定

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Isolation, Culture and Analysis of Mouse Mammary Epithelial Cells

Limited understanding of the cell biology of the breast and breast cancer hampers our ability to develop new therapeutic approaches. Mouse models of mammary gland development and tumourigenesis are key to developing new insights into the biology of both the normal and diseased tissues. Re ...

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Isolation and Culture of Mouse Pancreatic Islets for Ex Vivo Imaging Studies with Trappable or Recombinant Fluorescent Probes

The endocrine pancreas contains small clusters of 1,000–2,000 neuroendocrine cells termed islets of Langerhans. By secreting insulin, glucagon, or other hormones as circumstances dictate, islets play a central role in the control of glucose homeostasis in mammals. Islets are disper ...

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Isolation and Culture of Adult Mouse Hepatocytes

The liver performs a multitude of functions including the regulation of carbohydrate, fat, and protein metabolism, the detoxification of endo- and xenobiotics, and the synthesis and secretion of plasma proteins and bile. Isolated hepatocytes constitute a useful technique for study ...

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Isolation and Culture of Mouse Intestinal Cells

Complex cell signal transduction mechanisms regulate intestinal epithelial shape, polarity, motility, organelles, cell membrane components as well as physical and mechanical properties to influence alimentary digestion, absorption, secretion, detoxification and f ...

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Derivation of Primary Choroid Plexus Epithelial Cells from the Mouse

Choroid plexus epithelial cells form an integral and important part of the barrier between blood and cerebrospinal fluid. Culture of choroid plexus epithelium in vitro has been achieved from several mammalian species and this provides opportunities for the study of choroid plexus dev ...

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The Preparation of Primary Cortical Neuron Cultures and a Practical Application Using Immunofluorescent Cytochemistry

Traditionally, cultures of primary cortical neurons are prepared from embryonic animals because at prenatal stages neurons have not yet developed extensive axonal and dendritic arbors and are not highly innervated, thus rendering the cells less susceptible to damage during disso ...

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The Culture of Mouse Embryonic Stem Cells and Formation of Embryoid Bodies

Embryonic stem (ES) cells are pluripotent cells isolated from the inner cell mass of the pre-implantation blastocyst. They have the capacity to undergo indefinite rounds of self-renewing cell division and differentiate into all the cell lineages of the developing embryo. In suspension ...

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Isolation and Generation of Neurosphere Cultures from Embryonic and Adult Mouse Brain

Neural stem cells are defined as cells that either gives rise to or derives from the cells of the central nervous system and have the unique properties of stem cells, i.e. self-renewal and multipotentiality. One of the widely used methods of expanding neural stem cells under culture conditions is ba ...

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Cell Culture of Primary Cerebellar Granule Cells

Cerebellar granule cells are often used as a model system for the study of neuronal development, function and pathology, including the analysis of activity-dependent survival/apoptosis of neurons and the mechanisms of neuroprotection. Cerebellar granule cells are generated po ...

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Derivation of Primary Mouse Embryonic Fibroblast (PMEF) Cultures

Primary mouse embryonic fibroblasts (PMEFs) have a number of properties that make them an attractive cell culture model. Relative to other primary explant cultures they are easy to establish and maintain, proliferate rapidly and, as a result, large numbers of cells can be produced from a single ...

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Embryonic Skeletal Muscle Microexplant Culture and Isolation of Skeletal Muscle Stem Cells

Cultured embryonic and adult skeletal muscle cells have a number of different uses. The microdissected explant technique described in this chapter is a robust and reliable method for isolating relatively large numbers of proliferative skeletal muscle cells from juvenile, adult or em ...

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The Embryonic Kidney: Isolation, Organ Culture, Immunostaining and RNA Interference

Embryonic mouse kidneys develop well in organ culture. This, coupled with the fact that renal organogenesis includes a range of developmental processes, has made cultured kidney rudiments a popular model for the study of organogenesis. Although cultured kidneys do not replicate every e ...

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Explant Culture of Mouse Embryonic Whole Lung, Isolated Epithelium, or Mesenchyme Under Chemically Defined Conditions as a System to Evaluate the Mole

Lung primordial specification as well as branching morphogenesis, and the formation of various pulmonary cell lineages, requires a specific interaction of the lung endoderm with its surrounding mesenchyme and mesothelium. Lung mesenchyme has been shown to be the source of inductive ...

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Isolation, Culture, and Characterisation of Mouse Embryonic Oesophagus and Intestine

The gastrointestinal tract of vertebrates is lined by epithelium that develops from the endodermal germ layer. The oesophagus and intestine form part of the gastrointestinal tract and studying the normal development of both tissues is difficult due to lack of suitable in vitro model syst ...

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Isolation and Culture of Embryonic Pancreas and Liver

Culturing embryonic tissue in an in vitro setting offers the unique ability to manipulate the external medium and therefore to investigate the pathways involved in regulating normal organogenesis as well as providing models for developmental disorders. Here we describe a system for t ...

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Isolation and Culture of Mouse Satellite Cells

Muscle tissue culture provides a system for studying the growth and differentiation of muscle cells in a controlled environment. In mature muscle tissue, terminally differentiated myocytes form multinucleate syncytia in which structural and regulatory genes are expressed and t ...

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Analysis of Cardiac Myocyte Biology in Transgenic Mice: A Protocol for Preparation of Neonatal Mouse Cardiac Myocyte Cultures

We describe a method of isolating and maintaining primary cultures of mouse neonatal cardiac myocytes (NCM). This is derived from the well-established procedure for making NCM cultures from rat neonates by sequential digestion of rat ventricular myocardial pieces using a collagena ...

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Cell Surface Marker and Cell Cycle Analysis, Hedgehog Signaling, and Flow Cytometry

Detailed cytological analysis of cells undergoing differentiation often reveals clues to the regulation of multiple cell features. The Hedgehog (Hh) signaling cascade is a master regulator of cell fate during differentiation and is implicated in the development of some neoplasia ...

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Efficient Manipulation of Hedgehog/GLI Signaling Using Retroviral Expression Systems

Efficient manipulation of Hedgehog (HH)/GLI signaling activity is crucial to the analysis of molecular events underlying HH/GLI-regulated cell fate determination and tumor growth. In this article, we describe the use of retroviral expression systems as a valuable tool to activate or r ...

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Isolation of Rat Telencephalic Neural Explants to Assay Shh GABAergic Interneuron Differentiation-Inducing Activity

Multiple assays for Shh activity using cell lines, primary cultures, and explanted tissue have been described. We first described the use of E11.5 rat dorsal telencephalic explants to assay Shh ventralizing and differentiation-inducing activity in Kohtz et al. (1). Using this assay, we sub ...

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