1Regenerative Medicine Institute, Cedars-Sinai Medical Center
This protocol describes a novel mechanical chopping method that allows the expansion of spherical neural stem and progenitor cell aggregates without dissociation to a single cell suspension. Maintaining cell/cell contact allows rapid and stable growth for over 40 passages.
Published June 15, 2014. Keywords: Neuroscience, neural progenitor cell, neural precursor cell, neural stem cell, passaging, neurosphere, chopping, stem cell, neuroscience, suspension culture, good manufacturing practice, GMP
1NIH Stem Cell Unit, National Institute of Neurological Disorders and Stroke, National Institutes of Health, 2Craniofacial and Skeletal Diseases Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health
Here, we present human pluripotent stem cell (hPSC) culture protocols, based on non-colony type monolayer (NCM) growth of dissociated single cells. This new method, utilizing Rho-associated kinase inhibitors or the laminin isoform 521 (LN-521), is suitable for producing large amounts of homogeneous hPSCs, genetic manipulation, and drug discovery.
Published July 24, 2014. Keywords: Stem Cell Biology, Pluripotent stem cells, human embryonic stem cells, induced pluripotent stem cells, cell culture, non-colony type monolayer, single cell, plating efficiency, Rho-associated kinase, Y-27632, transfection, transduction
1Humanitas Clinical and Research Center, Italy, 2Institute of Genetic and Biomedical Research (IRGB), National Research Council (CNR)
Pluripotent stem cells, either embryonic or induced pluripotent stem (iPS) cells, constitute a valuable source of human differentiated cells, including cardiomyocytes. Here, we will focus on cardiac induction of iPS cells, showing how to use them to obtain functional human cardiomyocytes through an embryoid bodies-based protocol.
Published June 28, 2013. Keywords: Stem Cell Biology, Developmental Biology, Molecular Biology, Cellular Biology, Medicine, Bioengineering, Biomedical Engineering, Genetics, Cardiology, Stem Cell Research, Cardiovascular Diseases, Human cardiomyocytes, iPS cells, induced pluripotent stem cells, stem cells, cardiac differentiation, disease modeling, embryoid bodies, cell lines, cell culture
JoVE Developmental Biology
1Stanford Cardiovascular Institute, Stanford University School of Medicine, 2Institute of Stem Cell Biology and Regenerative Medicine, Cardiovascular Medicine Division, Department of Medicine, Child Health Research Institute, Stanford University School of Medicine
Here, we describe a robust protocol for human cardiomyocyte derivation that combines small molecule-modulated cardiac differentiation and glucose deprivation-mediated cardiomyocyte purification, enabling production of purified cardiomyocytes for the purposes of cardiovascular disease modeling and drug screening.
Published March 18, 2015. Keywords: Developmental Biology, Human induced pluripotent stem cells, cardiac differentiation, small molecule compounds, cardiomyocytes, glucose starvation
1Department of Cell and Molecular Physiology, Loyola University Chicago, 2Department of Biomedical Engineering, University of Alabama at Birmingham, 3Department of Bioengineering, Clemson University
This article describes the technique used to perform dual channel optical mapping in cultured HL-1 atrial cell monolayers. This unique protocol allows the simultaneous visualization of both calcium (Ca) and voltage (Vm) activity in the same area for the detailed detection and analysis of electrophysiological properties of culture monolayers.
Published March 23, 2015. Keywords: Cellular Biology, Cellular optical mapping, cultured atrial monolayer, action potential, calcium transient, voltage-sensitive dye, calcium dye
1Department of Biology, University of Rochester
This article describes GFP-based fluorescence in vivo assays that separately quantify homologous recombination and nonhomologous end joining in mammalian cells.
Published September 8, 2010. Keywords: Cellular Biology, DNA repair, HR, NHEJ, mammalian cells
1Molecular Pathology Laboratory Network, Inc
As cells reach confluency, they must be subcultured or passaged. This video will demonstrate a procedure for subculturing both adherent and suspension cells.
Published June 12, 2008. Keywords: Basic Protocols, Current Protocols Wiley, Cell Culture, Cell Passaging, Trypsinizing Cells, Adherent Cells, Suspension Cells