Developmental Biology

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    Protocols in Current Issue
    RNA Sequencing of Single Myofibers from Mus musculus
    Authors:  Darren M. Blackburn, Felicia Lazure and Vahab D. Soleimani, date: 02/20/2020, view: 166, Q&A: 0
    Whole transcriptome analysis is a key method in biology that allows researchers to determine the effect a condition has on gene regulation. One difficulty in RNA sequencing of muscle is that traditional methods are performed on the whole muscle, but ...
    RNA Sequencing of Single Myofibers from Mus musculus
    Authors:  Darren M. Blackburn, Felicia Lazure and Vahab D. Soleimani, date: 02/20/2020, view: 166, Q&A: 0
    [Abstract] Whole transcriptome analysis is a key method in biology that allows researchers to determine the effect a condition has on gene regulation. One difficulty in RNA sequencing of muscle is that traditional methods are performed on the whole muscle, but this captures non-myogenic cells that are part of the muscle. In order to analyze only the ...
    Mouse Embryonic Tooth Germ Dissection and Ex vivo Culture Protocol
    Authors:  Xue Han, Keigo Yoshizaki, Tian Tian, Kanako Miyazaki, Ichiro Takahashi and Satoshi Fukumoto, date: 02/05/2020, view: 162, Q&A: 0
    [Abstract] A tooth germ ex vivo organ culture allows visualization of its development in different stages, thus enabling investigation of the molecular mechanisms of regulatory factors. Tooth germs can be rapidly dissected from E13 mouse embryos and placed on cell culture inserts for observation of subsequent tooth germ development in a ...
    Isolation of Stem Cells, Endothelial Cells and Pericytes from Human Infantile Hemangioma
    Authors:  Lan Huang and Joyce Bischoff, date: 01/20/2020, view: 291, Q&A: 0
    [Abstract] Infantile hemangioma (IH) is a vascular tumor noted for its excessive blood vessel formation during infancy, glucose-transporter-1 (GLUT1)-positive staining of the blood vessels, and its slow spontaneous involution over several years in early childhood. For most children, IH poses no serious threat because it will eventually involute, but a subset ...
    RNA Interactome Identification via RNA-BioID in Mouse Embryonic Fibroblasts
    Authors:  Joyita Mukherjee, Mirita Franz-Wachtel, Boris Maček and Ralf-Peter Jansen, date: 01/05/2020, view: 394, Q&A: 0
    [Abstract] Cytoplasmic localization of mRNAs is common to all organisms and serves the spatial expression of genes. Cis-acting RNA signals (mostly found in the mRNA’s 3'-UTR), called zipcodes recruit trans-acting RNA-binding proteins that facilitate the localization of the mRNA. UV-cross-linking or affinity purification has been applied to identify such ...
    In vitro Differentiation of Human iPSC-derived Retinal Pigment Epithelium Cells (iPSC-RPE)
    Authors:  Agnieszka D’Antonio-Chronowska, Matteo D’Antonio and Kelly A. Frazer, date: 12/20/2019, view: 753, Q&A: 0
    [Abstract] Induced Pluripotent Stem Cells (iPSCs) serve as an excellent model system for studying the molecular underpinnings of tissue development. Human iPSC-derived retinal pigment epithelium (iPSC-RPE) cells have fetal-like molecular profiles. Hence, biobanks like iPSCORE, which contain iPSCs generated from hundreds of individuals, are an invaluable ...
    Mitochondrial Respiratory Measurements in Patient-derived Fibroblasts
    Authors:  Prashant Mishra, Ting Zhang, Ming Guo and David Chan, date: 12/05/2019, view: 481, Q&A: 0
    [Abstract] Mitochondrial dysfunction is associated with a number of human diseases. As an example, we recently established in vivo Drosophila models of IBMPFD (Inclusion body myopathy, Paget disease, and frontotemporal dementia), and uncovered that human disease mutations of the p97/VCP (Valosin Containing Protein) gene behave as ...
    Measurement of Mitotic Spindle Angle and Mitotic Cell Distance in Fixed Tissue of Drosophila Larval Brains
    Authors:  Maribel Franco and Ana Carmena, date: 11/20/2019, view: 445, Q&A: 0
    [Abstract] The positioning and the cleavage plane orientation of mitotic cells in pseudostratified epithelia (PSE) must be tightly regulated since failures in any of these processes might have fatal consequences during development. Here we present a simple method to determine the spindle orientation as well as the positioning of neuroepithelial mitotic cells ...
    Isolation and Culture of Murine Hepatic Stellate Cells
    Authors:  Rucha V. Modak and Dietmar M. Zaiss, date: 11/05/2019, view: 1062, Q&A: 0
    [Abstract] Hepatic stellate cells (HSCs), alternatively known as liver pericytes, can differentiate into myofibroblasts and secrete extra-cellular matrix components, thereby promoting wound healing and fibrosis. Studying HSCs can provide insights into the pathological mechanisms governing these processes. HSC isolation methods typically comprise of enzymatic ...
    Differentiation of Mouse Embryonic Stem Cells to Neuronal Cells Using Hanging Droplets and Retinoic Acid
    Authors:  Jeroen Witteveldt and Sara Macias, date: 11/05/2019, view: 596, Q&A: 0
    [Abstract] Controlled differentiation of embryonic stem cells is an essential tool in stem cell research. In this protocol, we describe a simple differentiation protocol involving the induction of embryoid body formation in mouse embryonic stem cells (mESC) using hanging droplets, followed by differentiation into a neuronal lineage.
    Cell Wall Compositional Analysis of Rice Culms
    Authors:  Lanjun Zhang, Baocai Zhang and Yihua Zhou, date: 10/20/2019, view: 648, Q&A: 0
    [Abstract] The plant cell wall is a complicated network that is mainly constituted of polysaccharides, such as cellulose, hemicellulose and pectin. Many noncellulosic polysaccharides are further acetylated, which confers these polymers flexible physicochemical properties. Due to the significance of cell wall in plant growth and development, the analytic ...



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