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  JoVE Biology

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  JoVE Medicine

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October, 2006
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 JoVE Immunology and Infection

Quantitative In vitro Assay to Measure Neutrophil Adhesion to Activated Primary Human Microvascular Endothelial Cells under Static Conditions

1Department of Anesthesia and Perioperative Care, University of California, San Francisco, 2Graduate Program in Biomedical Sciences, University of California, San Francisco

JoVE 50677

Neutrophil adherence to the activated endothelium at sites of infection is an integral component of the host's inflammatory response. Described in this report is a neutrophil binding assay that allows for the in vitro quantitation of primary human neutrophil binding to endothelial cells activated by inflammatory mediators under static conditions.

 JoVE Biology

Endothelial Cell Tube Formation Assay for the In Vitro Study of Angiogenesis

1Department of Biology, American University, 2Laboratory of Cancer Biology and Genetics, National Cancer Institute, NIH

JoVE 51312

The tube formation assay is a fast, quantifiable method for measuring in vitro angiogenesis. Endothelial cells are combined with conditioned media and plated on basement membrane extract. Tube formation occurs within hours and newly formed tubules easily quantified.

 JoVE Immunology and Infection

Expansion, Purification, and Functional Assessment of Human Peripheral Blood NK Cells

1Division of Pediatrics, MD Anderson Cancer Center - University of Texas

JoVE 2540

Here we describe a method to efficiently expand and purify large numbers of human NK cells and assess their function.

 JoVE Medicine

Stem Cell Transplantation in an in vitro Simulated Ischemia/Reperfusion Model

1Institute of Human Physiology and Clinical Experimental Research, Semmelweis University

JoVE 3575

We demonstrate how to set up an in vitro ischemia/reperfusion model and how to evaluate the effect of stem cell therapy on postischemic cardiac cells.

 JoVE Bioengineering

Tissue Engineering: Construction of a Multicellular 3D Scaffold for the Delivery of Layered Cell Sheets

1School of Engineering, University of California, Merced

JoVE 51044

For creation of highly organized structures of complex tissue, one must assemble multiple material and cell types into an integrated composite. This combinatorial design incorporates organ-specific layered cell sheets with two distinct biologically-derived materials containing a strong fibrous matrix base, and endothelial cells for enhancing new vessels formation.

 JoVE Medicine

Modeling and Imaging 3-Dimensional Collective Cell Invasion

1Strathclyde Institute for Pharmacy and Biomedical Sciences, University of Strathclyde, 2The Beatson Institute for Cancer Research

JoVE 3525

Models of tumor cell invasion into three-dimensional extracellular matrix better reflect the in vivo situation than two-dimensional motility assays. Using matrix invasion assays combined with confocal imaging of fluorescently-labeled cells, detailed information on invasion modes and the distinct contributions of leading versus following cells can be obtained.

 JoVE Medicine

Confocal Time Lapse Imaging as an Efficient Method for the Cytocompatibility Evaluation of Dental Composites

1Laboratoire des Multimatériaux et Interfaces, UMR CNRS 5615, Université Lyon1, 2UFR d'Odontologie, Université Lyon1; Service de Consultations et de Traitements Dentaires, Hospices Civils de Lyon, 3UFR d'Odontologie, Université Paris Diderot; Service d'Odontologie, APHP, Hôpital Rothschild

JoVE 51949

The most studied aspect of the biocompatibility of dental composites is their cytotoxicity 3D CLSM time lapse imaging combined with fluorescent signal quantification allows sensitive evaluation of the cell fate over time. Utilizing this method could be an efficient tool to assess the cytocompatibility of dental composites.

 JoVE Immunology and Infection

Visualization of Bacterial Toxin Induced Responses Using Live Cell Fluorescence Microscopy

1Department of Immunology, University of Pittsburgh School of Medicine, 2Department of Cell Biology and Physiology, University of Pittsburgh School of Medicine

JoVE 4227

Methods for purifying the cholesterol binding toxin streptolysin O from recombinant E. coli and visualization of toxin binding to live eukaryotic cells are described. Localized delivery of toxin induces rapid and complex changes in targeted cells revealing novel aspects of toxin biology.

 JoVE Bioengineering

High efficiency, Site-specific Transfection of Adherent Cells with siRNA Using Microelectrode Arrays (MEA)

1School of Biological and Health Systems Engineering, Arizona State University

JoVE 4415

The article details the protocol for site-specific transfection of scrambled sequence of siRNA in an adherent mammalian cell culture using a microelectrode array (MEA).

 JoVE Chemistry

Microwave-assisted Functionalization of Poly(ethylene glycol) and On-resin Peptides for Use in Chain Polymerizations and Hydrogel Formation

1Department of Biomedical Engineering, University of Rochester, 2Department of Chemical Engineering, University of Rochester, 3Center for Musculoskeletal Research, University of Rochester Medical Center

JoVE 50890

This video will illustrate a rapid, efficient method to methacrylate poly(ethylene glycol), enabling chain polymerizations and hydrogel synthesis. It will demonstrate how to similarly introduce methacrylamide functionalities into peptides, detail common analytical methods to assess functionalization efficiency, provide suggestions for troubleshooting and advanced modifications, and demonstrate typical hydrogel characterization techniques.

 JoVE Bioengineering

Multi-Scale Modification of Metallic Implants With Pore Gradients, Polyelectrolytes and Their Indirect Monitoring In vivo

1Biomatériaux et Bioingénieriee, INSERM, 2Service Oto-Rhino-Laryngologie, Hôpitaux Universitaires de Strasbourg, 3Faculté de Chirurgie Dentaire, Université de Strasbourg

JoVE 50533

In this video, we will demonstrate modification techniques for porous metallic implants to improve their functionality and to control cell migration. Techniques include development of pore gradients to control cell movement in 3D and production of basement membrane mimics to control cell movement in 2-D. Also, a HPLC-based method for monitoring implant integration in-vivo via analysis of blood proteins is described.

 JoVE Bioengineering

Encapsulation of Cardiomyocytes in a Fibrin Hydrogel for Cardiac Tissue Engineering

1Department of Biomedical Engineering, Tufts University

JoVE 3251

We describe the isolation of neonatal cardiomyocytes and the preparation of the cells for encapsulation in fibrin hydrogel constructs for tissue engineering. We describe methods for analyzing the tissue engineered myocardium after the culture period including active force generated upon electrical stimulation and cell viability and immunohistological staining.

 JoVE Immunology and Infection

Isolation of Human Monocytes by Double Gradient Centrifugation and Their Differentiation to Macrophages in Teflon-coated Cell Culture Bags

1Department of Hematology and Oncology, University Medical Center Göttingen, 2Microbial Interface Biology Group, Research Center Borstel

JoVE 51554

We present a simple and efficient protocol for the generation of human macrophages. Buffy coats are processed by double density gradient centrifugation and isolated monocytes are then differentiated to macrophages in Teflon-coated cell culture bags. This maximizes macrophage yields and facilitates cell harvesting for subsequent experiments.

 JoVE Biology

Platelet Adhesion and Aggregation Under Flow using Microfluidic Flow Cells

1Fluxion Biosciences, Inc.

JoVE 1644

The platelet adhesion cascade takes place in the presence of shear flow, a factor not accounted for in conventional (static) well-plate assays. This article reports on a platelet-aggregation assay utilizing a microfluidic well-plate format to emulate physiological shear flow conditions.

 JoVE Bioengineering

Fabrication of Micropatterned Hydrogels for Neural Culture Systems using Dynamic Mask Projection Photolithography

1Biomedical Engineering, Tulane University

JoVE 2636

Simple techniques are described for the rapid production of microfabricated neural culture systems using a digital micromirror device for dynamic mask projection lithography on regular cell culture substrates. These culture systems may be more representative of natural biological architecture, and the techniques described could be adapted for numerous applications.

 JoVE Biology

Cellular Encapsulation in 3D Hydrogels for Tissue Engineering

1Department of Bioengineering, University of Pennsylvania, 2Department of Bioengineering, University of Pennsylvania-School of Medicine

JoVE 1590

We present protocols for the 3-dimensional (3D) encapsulation of cells within synthetic hydrogels. The encapsulation procedure is outlined for two commonly used methods of crosslinking (michael-type addition and light-initiated free radical mechanisms), as well as a number of techniques for assessing encapsulated cell behavior.

 JoVE Medicine

Label-free Isolation and Enrichment of Cells Through Contactless Dielectrophoresis

1School of Biomedical Engineering and Science, Virginia Tech, 2Department of Engineering Science and Mechanics, Virginia Tech

JoVE 50634

Contactless dielectrophoresis (cDEP) achieves sorting and enrichment of particles via their intrinsic dielectric properties. Fluidic electrode channels replace metallic electrodes traditional to DEP, suiting cDEP to non-damaging sterile characterization and sorting of biological particles. We demonstrate how to prepare a cDEP microdevice and conduct cell characterization and sorting experiments.

 JoVE Bioengineering

Generation and Recovery of β-cell Spheroids From Step-growth PEG-peptide Hydrogels

1Department of Biomedical Engineering, Purdue School of Engineering and Technology, Indiana University - Purdue University at Indianapolis

JoVE 50081

The following protocol provides techniques for encapsulating pancreatic β-cells in step-growth PEG-peptide hydrogels formed by thiol-ene photo-click reactions. This material platform not only offers a cytocompatible microenvironment for cell encapsulation, but also permits user-controlled rapid recovery of cell structures formed within the hydrogels.

 JoVE Biology

Induction and Analysis of Epithelial to Mesenchymal Transition

1Antibody Development Department, R&D Systems, Inc., 2Stem Cells and Developmental Biology Department, R&D Systems, Inc.

JoVE 50478

A straightforward method is described for the induction of epithelial to mesenchymal transition (EMT) in a variety of cell types. Methods for the analysis of cells in EMT states by immunocytochemistry are included.

 JoVE Bioengineering

Robotic Production of Cancer Cell Spheroids with an Aqueous Two-phase System for Drug Testing

1Department of Biomedical Engineering, The University of Akron

JoVE 52754

A protocol for robotic printing of cancer cell spheroids in a high throughput 96-well plate format using an aqueous two-phase system is presented.

 JoVE Biology

A Functional Assay for Gap Junctional Examination; Electroporation of Adherent Cells on Indium-Tin Oxide

1Department of Microbiology and Immunology and Department of Pathology, Queen's University, 2Ask Science Products Inc.

JoVE 51710

This presentation demonstrates a method whereby electroporation of adherent, cultured cells is used for the study of intercellular, junctional communication, while the cells grow on a slide coated with conductive and transparent indium-tin oxide.

 JoVE Bioengineering

Constructing a Collagen Hydrogel for the Delivery of Stem Cell-loaded Chitosan Microspheres

1Department of Regenerative Medicine, United States Army Institute of Surgical Research

JoVE 3624

A major hurdle in current stem cell therapies is determining the most effective method to deliver these cells to host tissues. Here, we describe a chitosan-based delivery method that is efficient and simple in approach, while allowing adipose-derived stem cells to maintain their multipotency.

 JoVE Bioengineering

Preparation of Intact Bovine Tail Intervertebral Discs for Organ Culture

1ARTORG Center for Biomedical Engineering, University of Bern

JoVE 3490

This protocol illustrates a harvesting technique for coccygeal bovine intervertebral discs for organ culture for in vitro organ culture.

 JoVE Biology

Title Cell Encapsulation by Droplets

1Bio-Acoustic-MEMS Laboratory in Medicine (BAMM), HST-Center for Bioengineering, Brigham and Women's, Harvard Medical School, 2Bio-Acoustic-MEMS Laboratory in Medicine (BAMM), HST-Center for Bioengineering, Brigham and Women's Hospital, 3Brigham and Women's Hospital, Harvard Medical School, 4Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology; Center for Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital

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