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Assays of Lipolytic Enzymes

Free fatty acids (FAs) derived from adipose tissue (AT) are, quantitatively, the most important fuel in mammals, and provide more than half of the caloric needs during periods when dietary substrates are lacking. Hormone-sensitive lipase (HSL) catalyzes the rate-limiting step in the lipo ...

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Assays of Lipogenic Enzymes

The deposition of fat in adipose tissue (AT) is an important aspect of energy metabolism. AT fat originates either from ingested fatty acids (FAs) or from de novo synthesis. The synthesis of FAs is active in several tissues (liver, mammary gland, lung, intestine, brown and white ATs); however, in roden ...

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Embryonic Stem Cells as a Model for Studying Osteoclast Lineage Development

Osteoclasts are cells specialized to resorb bone (1,2). They originate from hematopoietic progenitors (3,4), which are widely distributed throughout the body. Osteoclast progenitors differentiate into cells that are positive for calcitonin receptor and tartrate-resistant ...

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Differentiation of Embryonic Stem Cells as a Model to Study Gene Function During the Development of Adipose Cells

The white adipose tissue stores energy in the form of triglycerides in time of nutritional excess and releases free fatty acids during food deprivation. The adipose tissue mass is determined by the balance between energy intake and expenditure. Alterations of this steady state can lead to ove ...

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Embryonic Stem Cell Differentiation and the Vascular Lineage

The ability of mouse embryonic stem (ES) cells to undergo differentiation in vitro complements their ability to contribute to numerous tissues in vivo and provides a unique model system for aspects of early mammalian development. ES cells are differentiated in two major ways: (i) unmanipul ...

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Cardiomyocyte Enrichment in Differentiating ES Cell Cultures: Strategies and Applications

Advances in our understanding of cardiomyocyte cell biology have been dependent largely upon the ability to generate primary cultures from enzymatically dispersed fetal, neonatal, or adult hearts. Primary cardiomyocyte cultures recapitulate many of the physiologic and mole ...

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Embryonic Stem Cells as a Model for the Physiological Analysis of the Cardiovascular System

Embryonic stem (ES) cells have the potential to proliferate infinitely in vitro in an undifferentiated and pluripotent state, thereby maintaining a relatively normal and stable karyotype even with continual passaging. Remarkably, in vivo, ES cells can be reincorporated into normal ...

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Isolation of Lineage-Restricted Neural Precursors from Cultured ES Cells

Embryonic stem (ES) cells are derived from undifferentiated cells present at the inner cell mass at the blastula stage of development (1,2). In the normal course of development, these cells give rise to the primitive ectoderm, which in turn gives rise to the ectoderm, mesoderm, and endoderm through ...

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Lineage Selection for Generation and Amplification of Neural Precursor Cells

Embryonic stem (ES) cells are derived from the epiblast of mouse blastocyst. They can repopulate all cell lineages in vivo and can differentiate into a wide variety of cell types in vitro during embryoid body (EB) formation (1). ES cells have been shown to generate both neurones and glial cells (2,3). Dur ...

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Selective Neural Induction from ES Cells by Stromal Cell-Derived Inducing Activity and Its Potential Therapeutic Application in Parkinson's Diseas

In vertebrate embryogenesis, the primordia of the nervous systems arise from uncommitted ectoderm during gastrulation. Spemann and Mangold demonstrated that the dorsal lip of the amphibian blastopore, which gives rise mainly to axial mesoderm, emanates inductive factors that di ...

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Epidermal Lineage

The epidermis is a stratified squamous epithelium that provides the protective layer of the skin. The mammalian epidermis is derived from embryonic ectoderm, and this layer eventually gives rise to a very early epithelial cell that further commits to become epidermal tissue (for review see ...

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ES Cell Differentiation Into the Hair Follicle Lineage In Vitro

Hair is arguably the most crucial skin appendage that shapes one’s self-image and acceptance in our society. However, despite this critical function, the precise regulatory steps governing the formation of hair follicles remains poorly understood. It is known, though, that hair follicle ...

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Methods for the Isolation and Maintenance of Murine Embryonic Stem Cells

Embryonic stem (ES) cells were first isolated in the 1980s by several independent groups. (1–4). These investigators recognized the pluripotential nature of ES cells to differentiate into cell types of all three primary germ lineages. Gossler et al. (5) described the ability and advantages of ...

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Embryonic Stem Cells as a Model for Studying Melanocyte Development

Melanocytes residing in the skin, inner ear, and uveal tract are derived from neural crest cells. They are highly dendritic, heavily pigmented, and are generally located in the epidermal basal cell layer of these areas, including hair follicles (1). The most convincing fact demonstrating the n ...

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ES Cell-Mediated Conditional Transgenesis

The use of mouse embryonic stem (ES) cells to alter the mouse genome has become a routine method to study gene function alongside classical transgenesis. Gene targeting by homologous recombination is the most widely used method of ES cell-mediated genome alterations. It allows the introduc ...

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Using Progenitor Cells and Gene Chips to Define Genetic Pathways

The zygote is a single cell, a fertilized egg, capable of giving rise to a complete organism. This is a truly remarkable transformation, with a single cell becoming trillions of cells or more, with a microscopic mass becoming in some cases tens or hundreds of kilograms, with a single amorphous cell making a ...

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Switching on Lineage Tracers Using Site-Specific Recombination

Methods to study the establishment and distribution of embryonic cell lineages have increased our understanding of the diverse events that comprise normal development. The resultant fate maps have also provided an important framework for systematically analyzing genotype-p ...

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Functional Genomics by Gene-Trapping in Embryonic Stem Cells

The pace of sequencing whole genomes by far exceeds our increase of knowledge on gene functions. Twenty-two genomes have been already completed and both the human and mouse genome are close to be completed as well. The number of sequences in the National Center for Biotechnology Information (NC ...

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From ES Cells to Mice: The Gene Trap Approach

The gene trap approach, represented in Fig. 1, is based on the use of mouse embryonic stem (ES) cells and a class of vectors containing a splice-acceptor site upstream of the reporter and resistance genes, β-galactosidase (β-gal, lacZ) and neomycin resistance (neo), respectively. Integration of ...

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Phage-Displayed Antibodies to Detect Cell Markers

Immunologic reagents such as antibodies can be valuable for identifying and isolating cells with particular characteristics. While antibodies derived from immunized animals are the most common immunologic reagents used for this purpose, “antibodies” selected by phage displ ...

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