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January 2015: This Month in JoVE - Introducing JoVE Developmental Biology

1, 2

1Department of Ophthalmology, Massachusetts Eye and Ear, 2JoVE Content Production

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    Summary

    Date Published: 1/06/2015, Issue 95; doi: 10.3791/5637

    Cite this Article

    Chao, W., Kolski-Andreaco, A. January 2015: This Month in JoVE - Introducing JoVE Developmental Biology. J. Vis. Exp. (95), e5637, doi:10.3791/5637 (2015).

    Abstract

    Here's a look at what's coming up in the January 2015 issue of JoVE: The Journal of Visualized Experiments.

    JoVE has been revolutionizing scientific publishing since 2006, when we released our first video articles in JoVE Biology. We've grown over the years-adding sections in Neuroscience, Immunology & Infection, Clinical & Translational Medicine, Bioengineering, Applied Physics, Behavior, Chemistry, and Environment.

    We are now pleased to introduce a new addition to the JoVE family: JoVE Development, which covers the entire field of developmental biology from the underlying genetic and epigenetic mechanisms to the growth and differentiation of single cells into organs and whole organisms.

    In many species, developmental processes offer clues about evolution. For example, the European lancelet (Branchiostoma lanceolatum) has many features of modern fish, but it doesn't have a backbone-so it's emerging as a useful model for studying the divergence of vertebrates from invertebrate ancestors. Hirsinger et al. have developed a method for visualizing embryonic development in the European lancelet. They inject oocytes with mRNAs that encode fluorescent proteins, and following fertilization, developmental processes can be visualized in vivo.

    Development also explores interactions between different cell types, and these interactions are fundamental for repairing and regenerating damaged or diseased tissues, like muscle. To study these interactions, Agley et al. take human skeletal muscle biopsies then purify and culture different cell types. They also characterize the cells using immunocytochemical methods that can be adapted to other cell types.

    One of the most exciting topics in developmental biology is the engineering of stem cells for experimental and clinical applications. This month we feature two stem cell protocols: Zielins et al. describe the isolation and purification of human adipose-derived stromal cells for bone engineering, and Lei et al. demonstrate how to differentiate embryonic stem cells into embryoid bodies, and then derive cardiac progenitor cells that further differentiate into cardiomyocytes and smooth muscle cells.

    Above all, developmental processes are fascinating to visualize, and JoVE Development features techniques for optimizing real-time imaging, such as this protocol for live-imaging of the Drosophila pupal eye. Using image-stabilization techniques, Hellerman et al. compensate for tissue movement and uneven topology to enhance the visualization of the developing Drosophila eye.

    You've just had a sneak peek of JoVE's new Development section in the January 2015 issue. Visit the website to see the full-length articles plus our other scientific sections in JoVE: The Journal of Visualized Experiments.

    Protocol

    Derivation of Cardiac Progenitor Cells from Embryonic Stem Cells

    Ieng Lam Lei1, Lei Bu2, Zhong Wang1

    1Cardiac Surgery, University of Michigan, 2Leon H Charney Division of Cardiology, New York University School of Medicine

    In this protocol, derivation of cardiac progenitor cells from both mouse and human embryonic stem cells will be illustrated. A major strategy in this protocol is to enrich cardiac progenitor cells with flow cytometry using fluorescent reporters engineered into the embryonic stem cell lines.

    Expression of Fluorescent Proteins in Branchiostoma lanceolatum by mRNA Injection into Unfertilized Oocytes

    Estelle Hirsinger1, João Emanuel Carvalho2, Christine Chevalier1,3, Georges Lutfalla5, Jean-François Nicolas1, Nadine Peyriéras4, Michael Schubert2

    1Département de Biologie du Développement et Cellules Souches, Institut Pasteur, 2Laboratoire de Biologie du Développement de Villefranche-sur-Mer (UMR7009 CNRS/UPMC Univ Paris 06), Sorbonne Universités, 3Equipe Epigenetic Control of Normal and Pathological Hematopoiesis, Centre de Recherche en Cancérologie de Marseille, 4Plateforme BioEmergences IBiSA FBI, CNRS-NED, Institut de Neurobiologie Alfred Fessard, 5Unité de Dynamique des Interactions Membranaires Normales et Pathologiques, CNRS UMR5235/DAA/cc107/Université Montpellier II

    We report here the robust and efficient expression of fluorescent proteins after mRNA injection into unfertilized oocytes of Branchiostoma lanceolatum. The development of the microinjection technique in this basal chordate will pave the way for far-reaching technical innovations in this emerging model system, including in vivo imaging and gene-specific manipulations.

    Isolation and Quantitative Immunocytochemical Characterization of Primary Myogenic Cells and Fibroblasts from Human Skeletal Muscle

    Chibeza C. Agley1,2, Anthea M. Rowlerson1, Cristiana P. Velloso1, Norman L. Lazarus1, Stephen D. R. Harridge1

    1Centre of Human and Aerospace Physiological Sciences, King‘s College London, 2Wellcome Trust-Medical Research Council, Cambridge Stem Cell Institute

    The main adherent cell types derived from human muscle are myogenic cells and fibroblasts. Here, cell populations are enriched using magnetic-activated cell sorting based on the CD56 antigen. Subsequent immunolabelling with specific antibodies and use of image analysis techniques allows quantification of cytoplasmic and nuclear characteristics in individual cells.

    Isolation and Enrichment of Human Adipose-derived Stromal Cells for Enhanced Osteogenesis

    Elizabeth R. Zielins*1, Ruth Tevlin*1, Michael S. Hu1, Michael T. Chung1, Adrian McArdle1, Kevin J. Paik1, David Atashroo1, Christopher R. Duldulao1, Anna Luan1, Kshemendra Senarath-Yapa1, Graham G. Walmsley1, Taylor Wearda1, Michael T. Longaker1,3, Derrick C. Wan1

    1Hagey Laboratory for Pediatric Regenerative Medicine, Department of Surgery, Division of Plastic and Reconstructive Surgery, Stanford University School of Medicine, 2Stem Cell and Biomaterials Engineering Laboratory, Department of Bioengineering, Stanford University School of Medicine, 3Institute for Stem Cell Biology and Regenerative Medicine, Stanford University

    The transcriptional heterogeneity within human adipose-derived stromal cells can be defined on the single cell level using cell surface markers and osteogenic genes. We describe a protocol utilizing flow cytometry for the isolation of cell subpopulations with increased osteogenic potential, which may be used to enhance craniofacial skeletal reconstruction.

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