<h2 class="jove_title"><a href="https://www.jove.com/video/52047">Derivation of Cardiac Progenitor Cells from Embryonic Stem Cells</a></h2>
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.
<h2 class="jove_title"><a href="https://www.jove.com/video/52042">Expression of Fluorescent Proteins in Branchiostoma lanceolatum by mRNA Injection into Unfertilized Oocytes</a></h2>
Estelle Hirsinger1, Jo&#227;o Emanuel Carvalho2, Christine Chevalier1,3, Georges Lutfalla5, Jean-Fran&#231;ois Nicolas1, Nadine Peyri&#233;ras4, Michael Schubert2
1D&#233;partement de Biologie du D&#233;veloppement et Cellules Souches, Institut Pasteur, 2Laboratoire de Biologie du D&#233;veloppement de Villefranche-sur-Mer (UMR7009 CNRS/UPMC Univ Paris 06), Sorbonne Universit&#233;s, 3Equipe Epigenetic Control of Normal and Pathological Hematopoiesis, Centre de Recherche en Canc&#233;rologie de Marseille, 4Plateforme BioEmergences IBiSA FBI, CNRS-NED, Institut de Neurobiologie Alfred Fessard, 5Unit&#233; de Dynamique des Interactions Membranaires Normales et Pathologiques, CNRS UMR5235/DAA/cc107/Universit&#233; 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.
<h2 class="jove_title"><a href="https://www.jove.com/video/52049"> Isolation and Quantitative Immunocytochemical Characterization of Primary Myogenic Cells and Fibroblasts from Human Skeletal Muscle </a></h2>
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&#8216;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. 
<h2 class="jove_title"><a href="https://www.jove.com/video/52181">Isolation and Enrichment of Human Adipose-derived Stromal Cells for Enhanced Osteogenesis</a></h2>
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.

january 2015: this month in jove - introducing jove developmental biology

Here's a look at what's coming up in the <a href="http://www.jove.com/archive/95">January 2015 issue</a> of <a href="http://www.jove.com/">JoVE: The Journal of Visualized Experiments</a>.
JoVE has been revolutionizing scientific publishing since 2006, when we released our first video articles in <a href="http://www.jove.com/biology">JoVE Biology</a>. We've grown over the years-adding sections in <a href="http://www.jove.com/neuro">Neuroscience</a>, <a href="http://www.jove.com/i2">Immunology &amp; Infection</a>, <a href="http://www.jove.com/ctm">Clinical &amp; Translational Medicine</a>, <a href="http://www.jove.com/bioengineering">Bioengineering</a>, <a href="http://www.jove.com/ap">Applied Physics</a>, <a href="http://www.jove.com/behavior">Behavior</a>, <a href="http://www.jove.com/chem">Chemistry</a>, and <a href="http://www.jove.com/enviro">Environment</a>.
We are now pleased to introduce a new addition to the JoVE family: <a href="http://www.jove.com/devbio">JoVE Development</a>, 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. <a href="http://www.jove.com/video/52042">Hirsinger et al.</a> 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, <a href="http://www.jove.com/video/52049">Agley et al.</a> 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: <a href="http://www.jove.com/video/52181">Zielins et al.</a> describe the isolation and purification of human adipose-derived stromal cells for bone engineering, and <a href="http://www.jove.com/video/52047">Lei et al.</a> 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 <a href="http://www.jove.com/devbio">JoVE Development</a> features techniques for optimizing real-time imaging, such as this protocol for live-imaging of the Drosophila pupal eye. Using image-stabilization techniques, <a href="http://www.jove.com/video/52120">Hellerman et al.</a> 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 <a href="http://www.jove.com/devbio">Development section</a> in the <a href="http://www.jove.com/archive/95">January 2015 issue</a>. Visit the website to see the full-length articles plus our other scientific sections in <a href="http://www.jove.com/">JoVE: The Journal of Visualized Experiments</a>.

Watch this Scientific Journal Video about January 2015: This Month in JoVE - Introducing JoVE Developmental Biology at JoVE.com

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

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 ...

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human pluripotent stem cell based developmental toxicity assays for chemical safety screening and systems biology data generation

Human Pluripotent Stem Cell Based Developmental Toxicity Assays for Chemical Safety Screening and Systems Biology Data Generation

The protocols describe two in vitro developmental toxicity test systems (UKK and UKN1) based on human embryonic stem cells and transcriptome studies. The test systems predict human developmental toxicity hazard, and may contribute to reduce animal studies, costs and the time required for chemical safety...

generation of 3d whole lung organoids from induced pluripotent stem cells for modeling lung developmental biology and disease

Generation of 3D Whole Lung Organoids from Induced Pluripotent Stem Cells for Modeling Lung Developmental Biology and Disease

The article describes step wise directed differentiation of induced pluripotent stem cells to three-dimensional whole lung organoids containing both proximal and distal epithelial lung cells along with...

introducing clicker training as a cognitive enrichment for laboratory mice

Introducing Clicker Training as a Cognitive Enrichment for Laboratory Mice

The development of new refinement strategies for laboratory mice is a challenging task that contributes towards fulfilling the 3R principle. This protocol introduces clicker training as a cognitive enrichment program for laboratory mice.

introducing shear stress in the study of bacterial adhesion

Introducing Shear Stress in the Study of Bacterial Adhesion

During the infection process, a key step is the adhesion of pathogens with host cells. In most instances this adhesion step occurs in the presence of mechanical stress generated by flowing liquid. We describe a technique that introduces shear stress as an important parameter in the study of bacterial...

intraoperative assessment of resection margins in oral cavity cancer: this is the way

Intraoperative Assessment of Resection Margins in Oral Cavity Cancer: This is the Way

The goal of this protocol is to provide a clear overview of specimen-driven intraoperative assessment of resection margins. It is encouraged to implement this protocol to improve patient care at other...

visual and microscopic evaluation of streptomyces developmental mutants

Visual and Microscopic Evaluation of Streptomyces Developmental Mutants

Here we present protocols for novice researchers to initiate phenotyping for the pharmacologically important bacterial genus...

Visual and Microscopic Evaluation of Streptomyces Developmental Mutants

measuring cardiac autonomic nervous system (ans) activity in toddlers - resting and developmental challenges

Measuring Cardiac Autonomic Nervous System ANS Activity in Toddlers - Resting and Developmental Challenges

We describe the methods for continuous monitoring of the autonomic nervous system under resting and challenge conditions with 18 month old children. Results revealed that this protocol can produce meaningful physiological responses in both branches of the autonomic nervous system and elicit significant individual variability in patterns of...

Measuring Cardiac Autonomic Nervous System (ANS) Activity in Toddlers - Resting and Developmental Challenges

tracking and quantifying developmental processes in c. elegans using open-source tools

Tracking and Quantifying Developmental Processes in C. elegans Using Open-source Tools

Here it is shown how to track and quantify developmental processes in C. elegans. The methods presented are based on open-source tools that can be easily implemented. It is demonstrated how to reconstruct 3D cell-shape models, how to manually track subcellular structures, and how to analyze cortical contractile...

Tracking and Quantifying Developmental Processes in C. elegans Using Open-source Tools

measuring composition of cd95 death-inducing signaling complex and processing of procaspase-8 in this complex

Measuring Composition of CD95 Death-Inducing Signaling Complex and Processing of Procaspase-8 in this Complex

Here, an experimental workflow is presented that enables the detection of caspase-8 processing directly at the death-inducing signaling complex (DISC) and determines the composition of this complex. This methodology has broad applications, from unraveling the molecular mechanisms of cell death pathways to the dynamic modeling of apoptosis...

in vitro model of fetal human vessel on-chip to study developmental mechanobiology

Author Spotlight: Studying hiPSC-Derived Endothelial Cells Cultured Under Fluidic-Mediated Mechanical Stimulation 

Described here is a simple workflow to differentiate endothelial cells from human pluripotent stem cells followed by a detailed protocol for their mechanical stimulation. This allows for the study of the developmental mechanobiology of endothelial cells. This approach is compatible with downstream assays of live cells collected from the culture chip after mechanical...

In Vitro Model of Fetal Human Vessel On-chip to Study Developmental Mechanobiology

application of aorta-gonad-mesonephros explant culture system in developmental hematopoiesis

Application of Aorta-gonad-mesonephros Explant Culture System in Developmental Hematopoiesis

This protocol describes using cultured Aorta-Gonad-Mesonephros for expression analyses, colony-forming units in the culture and spleen, and long-term reconstitution to determine the effect of regulatory factors and signaling pathways on hematopoietic stem cell development. This has been demonstrated as an effective system for studying hematopoietic stem cell biology and...

the use of chemostats in microbial systems biology

The Use of Chemostats in Microbial Systems Biology

Cell growth rate is a regulated process and a primary determinant of cell physiology. Continuous culturing using chemostats enables extrinsic control of cell growth rate by nutrient limitation facilitating the study of molecular networks that control cell growth and how those networks evolve to optimize cell...

involving individuals with developmental language disorder and their parents/carers in research priority setting

Involving Individuals with Developmental Language Disorder and Their Parents/Carers in Research Priority Setting

A protocol to enable individuals with developmental language disorder (DLD) and their parents/carers to meaningfully participate in a research priority setting exercise is established. The protocol includes a defined program of activities for data collection, and methods to incorporate this data into the broader research priority setting...

experience is instrumental in tuning a link between language and cognition: evidence from 6-  to 7- month-old infants' object categorization

Experience is Instrumental in Tuning a Link Between Language and Cognition: Evidence from 6-  to 7- Month-Old Infants' Object Categorization

At 3-4 months, listening to human and nonhuman primate vocalizations boosts infants&#39; cognition; by 6 months, only human vocalizations exert this cognitive advantage. We describe an exposure manipulation that reveals the powerful shaping role of experience as infants specify which sounds to link to cognition and which to tune...

windowing chicken eggs for developmental studies

Windowing Chicken Eggs for Developmental Studies

The ease of accessibility of the embryo has allowed for experiments to map cell fates using several approaches, including chick quail chimeras and focal dye labeling. In this article we demonstrate one egg preparation method that has been optimized for long survival...

using chicken embryo as a powerful tool in assessment of developmental cardiotoxicities

Using Chicken Embryo as a Powerful Tool in Assessment of Developmental Cardiotoxicities

Chicken embryos, as a classical developmental model, are used in our lab to assess developmental cardiotoxicities following exposure to various environmental contaminants. Exposure methods and morphological/functional assessment methods established are described in this manuscript.

a human ex vivo atherosclerotic plaque model to study lesion biology

A Human Ex Vivo Atherosclerotic Plaque Model to Study Lesion Biology

Atherosclerosis is a chronic inflammatory process. This manuscript illustrates an easy to use ex vivo model to investigate fresh carotid or coronary artery plaques. The ex vivo model allows for the investigation of potential substances on the inflammatory milieu in human atherosclerotic lesions and results can be analyzed by various...

A Human Ex Vivo Atherosclerotic Plaque Model to Study Lesion Biology

introducing point mutations into human pluripotent stem cells using seamless genome editing

Introducing Point Mutations into Human Pluripotent Stem Cells Using Seamless Genome Editing

Here, we describe a detailed method for seamless gene editing in human pluripotent stem cells using a piggyBac-based donor plasmid and the Cas9 nickase mutant. Two point mutations were introduced into exon 8 of the hepatocyte nuclear factor 4 alpha (HNF4&#945;) locus in human embryonic stem cells...

introducing a gene knockout directly into the amastigote stage of trypanosoma cruzi using the crispr/cas9 system

Introducing a Gene Knockout Directly Into the Amastigote Stage of Trypanosoma cruzi Using the CRISPR/Cas9 System

Here, we describe a protocol to introduce a gene knockout into the extracellular amastigote of Trypanosoma cruzi, using the CRISPR/Cas9 system. The growth phenotype can be followed up either by cell counting of axenic amastigote culture or by proliferation of intracellular amastigotes after host cell...

Introducing a Gene Knockout Directly Into the Amastigote Stage of Trypanosoma cruzi Using the CRISPR/Cas9 System

August 2012: This Month in JoVE

Traditional microscopy requires lens objectives to magnify specimens, and can involve numerous optical components like additional objectives, filters, and mirrors to refract and direct light to optical sensors. The August 2012 issue of JoVE (Journal of Visualized Experiments) is marked by the third publication from the Ozcan Lab (University of California, Los Angeles) on their lens free lens-free "on-chip" microscopy platform, which they have pioneered.
Back in 2009, the Ozcan lab published their first article, which demonstrated the use of a lens-free imaging platform that could image cells within whole blood. This imaging technique involves placing a sample directly on an optoelectric sensor array, like the CCD or CMOS chips found in the standard camcorder. Then, LED illumination from above the sample casts shadows on the chip, which are recorded as digital holograms that can be subjected to image processing. Not only does this lensless imaging system allow for an extremely large field of view (FOV of 0.6-8 cm2), the lack of bulky optical components makes it excellent for microfluidic applications. The Ozcan group demonstrates this in their follow-up 2011 JoVE article, in which fluorescently labeled white blood cells are imaged within a microfluidic chip.
This August in JoVE, the Ozcan group demonstrates further advancements to their lens-free microscopy platform by applying it to tomographic imaging of the nematode, C. elegans. By changing the angle of illumination, using a motorized rotation stage, the Ozcan group shows how multiple holographic slices may be generated for a sample-which, following processing, can be combined into tomograms. The Ozcan group confirms that their 3D reconstruction of the worm is successful by citing that the pharyngeal tube, a 5-micron-thick structure, is only visible in a single slice and in a 3D reconstructed cross section.
The compact and inexpensive nature of the lens-free imaging systems - designed and assembled by the Ozcan group - make them well suited for point-of-care diagnostic applications, or even cell phone based microscopy that can be performed in the field.
At the Ernest Gallo Clinic and Research Center at University of California, San Francisco JoVE learns how to visualize morphological changes in single cells using three-dimensional fluorescence microscopy and computer analysis. In this procedure, cells are given agonist, antagonist plus agonist, or no treatment, and prepared for immunofluorescence microscopy. Three-dimensional images are acquired with the help of a two-photon laser system, and the fluorescence data are processed on the computer using Imaris and Matlab.
To demonstrate the power of this technique, our authors quantify single-cell phenotypic changes in transfected HEK293 cells. Three-dimensional morphometric analysis shows dramatic changes in cells treated with agonist. These changes are not always apparent when using conventional 2D analysis. This technique allows morphological changes in single cells to be quantified and characterized in response to pharmacologic stimuli, offering a powerful new assessment tool to the arsenal of drug discovery research.
This month, in JoVE Applied Physics, we travel to South Carolina to visit the School of Materials Science and Engineering at Clemson University. Here, researchers show us how to construct lithium batteries for laboratory research.
The basic components of a lithium coin cell are the working electrode, the lithium electrolyte, and a lithium foil counter electrode. The different components of the coin cell are carefully assembled and sealed with a crimping machine. The finished coin cell can then be characterized in a battery tester. Using their homemade coin cells, our researchers can experiment with using different materials to make the cathodes of lithium coin cells.
This month in JoVE Immunology and Infection, we head to Basel, Switzerland to witness two techniques for diagnosing helminth infections. We also see how investigators perform the shuttle run test in children, in order to assess the effects of these infections on physical fitness.
Helminths are parasitic worms that can live inside humans and animals. Many species infect the intestinal tract, and may be transmitted through soil. Our authors demonstrate the Kato-Katz technique, which is used for diagnosing several common soil-transmitted helminths. They also demonstrate the Baermann technique, which uses light to stimulate the migration of certain types of phototactic helminths out of sample preparations. The various types of helminth species can subsequently be determined based on morphology.
Our authors then demonstrate how to set up and perform a 20-meter shuttle run test for children, and to measure the impact of helminth infections on physical fitness. This method can help determine the true burden of soil-transmitted helminth infections on overall health.
This brief summary highlights but a few notable video-articles that will be released in August. We'll also feature articles that demonstrate how to express genes in the midbrain of the chick embryo, study neutrophil transmigration in parallel flow chambers lined with endothelial cells, and precisely measure proteolytic degradation of the extracellular matrix.

Traditional microscopy requires lens objectives to magnify specimens, and can involve numerous optical components like additional objectives, filters, and mirrors to refract and direct light to optical sensors. The August 2012 issue of JoVE (Journal of Visualized Experiments) is marked by the third publication from the Ozcan Lab (University of California, Los Angeles) on their lens-free "on-chip" microscopy platform, which they have pioneered.

Traditional microscopy requires lens objectives to magnify specimens, and can involve numerous optical components like additional objectives, filters, and ...