1Center for Biomaterials and Tissue Engineering, Universitat Politècnica de València, 2Division of Biomedical Engineering, School of Engineering, University of Glasgow, 3Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN)
The following protocol describes the procedure to assemble sandwich-like cultures to be used as an intermediate stage between bi-dimensional (2D) and three-dimensional (3D) cellular environments. The engineered systems can have applications in microscopy, biomechanics, biochemistry and cell biology assays.
Published August 4, 2015. Keywords: Bioengineering, Sandwich culture, 3D culture, cell culture, physiological, cellular environment, bioengineering
1Woodruff School of Mechanical Engineering, Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, 2Coulter Department of Biomedical Engineering, Georgia Institute of Technology, 3Charles Perkins Centre, The University of Sydney, 4Institute of Biophysics, Laboratory of RNA Biology, Chinese Academy of Sciences, 5University of Chinese Academy of Sciences, 6School of Medicine and Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Zhejiang University
We describe a technique for concurrently measuring force-regulated single receptor-ligand binding kinetics and real-time imaging of calcium signaling in a single T lymphocyte.
Published August 4, 2015. Keywords: Bioengineering, single cell, single molecule, receptor-ligand binding, kinetics, fluorescence and force spectroscopy, adhesion, mechano-transduction, calcium
JoVE Developmental Biology
1Department of Biomedical Science, Graduate School of Medicine, Korea University
This protocol provides a simple and efficient way to propagate human pluripotent stem cells (hPSCs) using only conditioned media derived from the human placenta in a gelatin-coated dish without additional exogenous supplementation or hPSC-specific synthetic substrata.
Published August 3, 2015. Keywords: Developmental Biology, bFGF, Human embryonic stem cells, Human placenta chorionic villi, induced-pluripotent stem cells, Stem cell biology, Synthetic substrates
JoVE Developmental Biology
1Nephrology Division, Department of Medicine, Massachusetts General Hospital, 2Basic Sciences Division, Fred Hutchinson Cancer Research Center, 3Department of Genetics, Harvard Medical School
Here we present a reliable method to study adult kidney regeneration by inducing acute kidney injury by gentamicin injection. We show that injury is dependent on gentamicin dosage and environmental temperature using in situ hybridization to label lhx1a+ developing new nephrons.
Published August 3, 2015. Keywords: Developmental Biology, kidney, mesonephros, gentamicin, injection, zebrafish, injury, regeneration, Lhx1a, heat shock, adult
1Department of Ophthalmology, Massachusetts Eye and Ear, 2JoVE Content Production
Published August 2, 2015. Keywords: This Month in JoVE,
1Department of Genetics, Stanford University, 2Stanford Center for Genomics and Personalized Medicine, Stanford University
Using protein microarrays containing nearly the entire S. cerevisiae proteome is probed for rapid unbiased interrogation of thousands of protein-protein interactions in parallel. This method can be utilized for protein-small molecule, posttranslational modification, and other assays in high-throughput.
Published August 2, 2015. Keywords: Cellular Biology, Protein microarrays, kinase, yeast, protein-protein interactions
1Environmental Health Science and Research Bureau, Health Canada
The goal of this protocol is the detection of the DNA oxidation marker, 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dGuo) by HPLC-ED, in DNA from cultured cells or animal tissues.
Published August 1, 2015. Keywords: Chemistry, Oxidative Stress, DNA Damage, 8-oxo-7, 8-dihydro-2'-deoxyguanosine, 8-hydroxy-2'-deoxyguanosine, Xenobiotic Metabolism, Human Health
1Division of Drug Delivery and Tissue Engineering, University of Nottingham, 2Laboratory of Biophysics and Surface Analysis, School of Pharmacy, University of Nottingham, 3Division of Immunology and Allergy, School of Molecular Medical Sciences, University of Nottingham, 4Division of Respiratory Medicine, School of Clinical Sciences, University of Nottingham, 5NIHR Respiratory Biomedical Research Unit, University of Leicester, 6School of Sport, Exercise, and Health Sciences, Loughborough University
Advancements in biomaterial technologies enable the development of three-dimensional multi-cell-type constructs. We have developed electrospinning protocols to produce three individual scaffolds to culture the main structural cells of the airway to provide a 3D in vitro model of the airway bronchiole wall.
Published July 31, 2015. Keywords: Bioengineering, Electrospinning, 3D Cell Culture, Bioreactor, Airway, Tissue Engineering, In Vitro Model
1Graduate School of Medicine, Laboratory of Clinical Biotechnology, The University of Tokyo, 2Graduate School of Engineering, Department of Materials Engineering, The University of Tokyo, 3Graduate School of Engineering, Department of Bioengineering, The University of Tokyo
This protocol describes a cell transplantation system using genetically modified, injectable spheroids. Cell spheroids are cultured on micropatterned culture plates and recovered after gene introduction using polyplex nanomicelles. This system facilitates prolonged transgene expression from the transplanted cells in host animals while maintaining the innate function of the cells.…
Published July 31, 2015. Keywords: Bioengineering, Spheroid, Cell transplantation, Gene transfection, Non-viral carrier, Micropatterned plate, Genetic modification
JoVE Developmental Biology
1Department of Biological Sciences, University of Notre Dame
The zebrafish is an excellent model system for genetic and developmental studies. Bead implantation is a valuable tissue manipulation technique that can be used to interrogate developmental mechanisms by introducing alterations in local cellular environments. This protocol describes how to perform microbead implantation in the zebrafish embryo.
Published July 30, 2015. Keywords: Developmental Biology, zebrafish, embryo, bead implantation, tissue manipulation, development, organogenesis