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Three-Dimensional Culture for Expansion and Differentiation of Embryonic Stem Cells

Although embryonic stem (ES) cells have the regenerative capability of producing any tissue in our body, the development of efficient methods to generate a large number of ES cells with high purity has still remained a technical challenge before they can be used routinely in clinical trials. In t ...

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Scale-Up of Single CellInoculated Suspension Cultures of Human Embryonic Stem Cells

We have developed a simple yet highly reproducible technique of upscaling single cell–inoculated human embryonic stem cells (hESCs) in suspension cultures, using defined, nonconditioned media. Mass expansion of hESC was readily achieved by serially upscaling 2-ml static cultur ...

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Laboratory-Scale Purification of a Recombinant E-Cadherin-IgG Fc Fusion Protein That Provides a Cell Surface Matrix for Extended Culture and Efficient

The culture of human pluripotent stem cells under defined conditions most commonly relies on the use of Matrigel as a substrate upon which the cells remain in an undifferentiated state. Matrigel is a complex mixture of extracellular matrices and growth factors derived from mouse Engelbre ...

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Growth of Human Embryonic Stem Cells in Long-Term Hypoxia

Human embryonic stem cells (hESCs) hold a great promise for regenerative medicine and tissue engineering. In order to obtain uniform hESC cultures without spontaneous differentiation, which is of interest for basic investigations as well as the development of future therapeutic pr ...

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Feeder-Free Growth of Undifferentiated Human Embryonic Stem Cells

Conventionally, human embryonic stem cells (hESCs) are cultured on feeder cells. The most commonly used feeder cells are mouse embryonic fibroblasts. It is thought that the feeder cells provide an optimal microenvironment for the undifferentiated growth of hESCs by supplying curren ...

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Optimization of the Caco-2 Permeability Assay to Screen Drug Compounds for Intestinal Absorption and Efflux

In vitro permeability assays are a valuable tool for scientists during lead compound optimization. As a majority of discovery projects are focused on the development of orally bioavailable drugs, correlation of in vitro permeability data to in vivo absorption results is critical for und ...

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Studying Permeability in a Commonly Used Epithelial Cell Line: T84 Intestinal Epithelial Cells

The integrity, or barrier function, of the intestinal epithelium is of paramount importance in �maintaining good health. This is largely imparted by a single layer of epithelial cells linked by the transmembrane tight junction protein complex near their apical surface. Disruption of ep ...

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Analysis of Epithelial Cell Shedding and Gaps in the Intestinal Epithelium

The intestinal barrier is formed by a monolayer of columnar epithelial cells. This barrier is effectively maintained despite the high turnover of epithelial cells in the gut. Defects in the mechanism by which barrier function is maintained are believed to play a central role in the pathogenes ...

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Assessment of Intestinal Permeability in (Premature) Neonates by Sugar Absorption Tests

Infants born prematurely have an enhanced intestinal permeability compared to healthy term infants. This enhanced permeability might be a contributing factor in the development of Necrotising Enterocolitis. The assessment of intestinal permeability in premature neonates ...

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Assessment of Permeability Barriers to Macromolecules in the Rodent Endometrium at the Onset of Implantation

In rodents, embryo implantation is an invasive process, which begins with its attachment to the uterine wall and culminates in the formation of the definitive placenta several days later. It is critical that the endometrium provide a supportive environment for the implanting embryo duri ...

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Epidermal Permeability Barrier Measurement in Mammalian Skin

A defective skin epidermal permeability barrier (EPB) is responsible for a high mortality rate in premature infants and is an important risk factor in inflammatory skin diseases such as eczema. We report here fast and accurate methods for measurement of EPB in animal models or in human patients u ...

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Transcriptional Regulation of Epidermal Barrier Formation

The mammalian epidermis is a self-renewing stratified squamous epithelium. Its basal cell layer contains proliferating keratinocytes that exit the cell cycle when they move into the suprabasal compartment. These cells activate a gene differentiation program aimed at building a p ...

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In Vitro Human Skin Segmentation and Drug ConcentrationSkin Depth Profiles

Highly optimized methods for skin segmentation are provided using tape stripping in combination with infrared absorption measurements for stratum corneum (SC) and cryosectioning for deeper skin layers. Furthermore, an example is calculated for demonstration of the respecti ...

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In Vivo Optical Imaging of Ischemic BloodBrain Barrier Disruption

The blood–brain barrier (BBB) disruption following cerebral ischemia (stroke) contributes to the development of life-threatening brain edema. Recent studies suggested that the ischemic BBB disruption is not uniform throughout the affected brain region. The aim of this study was to e ...

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In Vivo Measurement of Glioma-Induced Vascular Permeability

The normal blood–brain barrier (BBB) consists of tight interendothelial cell junctions and adjacent astrocyte end feet separated by a basal lamina surrounding the endothelium. The interactions between the different cell types of BBB are disrupted in distinct patterns in the microe ...

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Evaluation of VEGF-Induced Vascular Permeability in Mice

Vascular endothelial growth factor (VEGF) is a potent inducer of angiogenesis and vascular leak involved in development, wound healing, tumor growth, macular degeneration, and ischemia. Studying the effects of VEGF in vitro is not always sufficient to approximate the complex in vivo res ...

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In Vitro and In Vivo Methods for Assessing FcRn-Mediated Reverse Transcytosis Across the BloodBrain Barrier

The neonatal Fc receptor, FcRn, mediates endocytic recycling pathway that prevents degradation of IgG and is expressed in most endothelial cells. The blood–brain barrier (BBB), formed by brain endothelial cells sealed with tight junctions, restricts transport of IgG from the blood to the ...

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Assessment of Permeability in Barrier Type of Endothelium in Brain Using Tracers: Evans Blue, Sodium Fluorescein, and Horseradish Peroxidase

Blood–brain barrier (BBB) constituted primarily by the capillary endothelial cells functions to maintain a constant environment for the brain, by preventing or slowing down the passage of a variety of blood-borne substances, such as serum proteins, chemical compounds, ions, and hormo ...

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Size-Selective and In Vitro Assessment of Inner Blood Retina Barrier Permeability

Assessment of tight junction integrity in vitro is fundamental when studying molecular processes that may be implicated in barrier dysfunction. At the blood brain and inner blood retina barrier (BBB and iBRB, respectively) adjacent endothelial cells lining the microvasculature h ...

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Permeability of Endothelial Barrier: Cell Culture and In Vivo Models

The methods for assessment of endothelial barrier permeability are vital tools of experimental biology. They allow us to measure permeability of endothelial monolayer in cell culture and in lung vessels or to determine formation of tissue edema resulting from increased permeabili ...

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