JoVE Bioengineering merges both physical and life sciences to understand and predict biological processes. Applying physical science tools to life science questions allow for the discovery of better technologies to measure, diagnose, and clinically treat disease.
1Department of Pharmaceutical Sciences, Mary & Dick Holland Regenerative Medicine Program, University of Nebraska Medical Center, 2Department of Orthopedic Surgery, Joan C. Edwards School of Medicine, Marshall University
Here, we present a protocol to fabricate electrospun nanofiber scaffolds with gradated organization of fibers and explore their applications in regulating cell morphology/orientation. Gradients with regard to physical and chemical properties of the nanofiber scaffolds offer a wide variety of applications in the biomedical field.
Published April 19, 2015. Keywords: Bioengineering, Electrospinning, Nanofiber scaffolds, Gradations, Stem cells, Tissue engineering, Nanoencapsulation
1School of Materials, The University of Manchester
This article describes a range of set-ups for seeding human mesenchymal stem cells onto materials, in this case electrospun yarns, that do not cover the base of standard culture well plates in order to maximize and quantify the number of cells that initially attach compared to the known seeding density.
Published April 10, 2015. Keywords: Bioengineering, Human mesenchymal stem cells, Electrospinning, Cell seeding, Cell attachment, Scaffold, Rotary vessel bioreactor, Cell number, Cell DNA assay, SEM, Low binding well plate, Poly(ε-caprolactone), Electrospun yarn
1Systems Biophysics Department, FOM Institute AMOLF, 2Mechanobiology Institute, National University of Singapore, 3Department of Biomedical Engineering, National University of Singapore
The mechanical properties and microstructure of the extracellular matrix strongly affect 3D migration of cells. An in vitro method to study the spatiotemporal cell migration behavior in biophysically variable environments, at both population and individual cell levels, is described.
Published April 3, 2015. Keywords: Bioengineering, cell migration, collagen, biomechanics, 3D cell culture, live-cell imaging, cancer invasion, metastasis, extracellular matrix, pore size, biopolymer, cytoskeleton, confocal microscopy
1Institute of Nanoengineering and Microsystems, National Tsing Hua University, 2Taichung Veterans General Hospital
This protocol details a method to isolate extracellular vesicles (EVs), small membranous particles released from cells, from as little as 10 μl serum samples. This approach circumvents the need for ultracentrifugation, requires only a few minutes of assay time, and enables the isolation of EVs from samples of limited volumes.
Published April 3, 2015. Keywords: Bioengineering, extracellular vesicles, exosomes, cellulose paper, microfluidics, paper ELISA, aqueous humor, chemical conjugation
1Department of Physiology & Bio-Physics, State University of New York Buffalo School of Medicine, 2Department of Pediatrics, State University of New York Buffalo School of Medicine, 3Department of Chemical and Biological Engineering, State University of New York Buffalo School of Engineering
A step-by-step protocol for the inter-positional placement of Tissue Engineered Vessels (TEVs) into the carotid artery of a sheep using end-to-end anastomosis and real-time digital assessment in vivo until animal sacrifice.
Published April 3, 2015. Keywords: Bioengineering, Vascular surgery, Tissue Engineered Vessel, Surgical Technique, Bio-Engineering, Vascular Grafts, Implantation, Sheep, Large animal model, Carotid Artery, Anastomosis
1Department of Orthopaedic Surgery, University of California Los Angeles, 2Department of Bioengineering, University of California Los Angeles
Animal models are important tools for the evaluation of tissue-engineered grafts. This paper presents the protocol for preparing an electrospun biodegradable polymer graft for use in anterior cruciate ligament tissue engineering, as well as a surgical protocol for implantation in a rat model.
Published March 26, 2015. Keywords: Bioengineering, Anterior cruciate ligament, tissue engineering, animal model, biodegradable scaffold, rat, knee
1Biomedical Engineering Department, Saint Louis University
Here, a method that enables quick, efficient, and inexpensive preparation of polyacrylamide gels in a multiwell plate format is described. The method does not require any specialized equipment and could be easily adopted by any research laboratory. It would be particularly useful in research focused on understanding stiffness-dependent cell responses.
Published March 25, 2015. Keywords: Bioengineering, Multiwell, substrate stiffness, drug screening, polyacrylamide, Young’s modulus, high-throughput
1Cardiovascular Division, King's College London BHF Centre
Here, we present a protocol to generate tissue engineered vessel grafts that are functional for grafting into mice by double seeding partially induced pluripotent stem cell (PiPSC) - derived smooth muscle cells and PiPSC - derived endothelial cells on a decellularized vessel scaffold bioreactor.
Published March 18, 2015. Keywords: Bioengineering, stem cells, partially induced pluripotent stem cells, tissue engineering, bioreactor, vascular differentiation, vessel graft, mouse models
1Division of Chemistry and Chemical Engineering, California Institute of Technology, 2Department of Molecular Medicine, Beckman Research Institute of the City of Hope
Gallium(III) 5,10,15-(tris)pentafluorophenylcorrole and its freebase analogue exhibit low micromolar cell cytotoxicity. This manuscript describes an RNA transcription reaction, imaging RNA with an ethidium bromide-stained gel, and quantifying RNA with UV-Vis spectroscopy, in order to assess transcription inhibition by corroles and demonstrates a straightforward method of evaluating anticancer candidate properties.
Published March 18, 2015. Keywords: Bioengineering, Corrole, RNA, transcription, inhibition, anti-cancer, DNA, binding, Actinomycin D, triptolide
1Bredesen Center, University of Tennessee, Knoxville, 2Center for Nanophase Materials Sciences, Oak Ridge National Laboratory, 3Department of Materials Science and Engineering, University of Tennessee, Knoxville
A microfabricated device with sealable femtoliter-volume reaction chambers is described. This report includes a protocol for sealing cell-free protein synthesis reactants inside these chambers for the purpose of understanding the role of crowding and confinement in gene expression.
Published March 11, 2015. Keywords: Bioengineering, Cell-free, synthetic biology, microfluidics, noise biology, soft lithography, femtoliter volumes