Developmental Biology

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    Protocols in Current Issue
    Isolation and Culture of Murine Hepatic Stellate Cells
    Authors:  Rucha V. Modak and Dietmar M. Zaiss, date: 11/05/2019, view: 195, Q&A: 0
    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 ...
    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: 92, Q&A: 0
    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) ...
    Isolation and Culture of Murine Hepatic Stellate Cells
    Authors:  Rucha V. Modak and Dietmar M. Zaiss, date: 11/05/2019, view: 195, 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: 92, 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: 292, 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 ...
    Immunohistochemical Identification of Muscle Fiber Types in Mice Tibialis Anterior Sections
    Authors:  Vanitha V. Rao and Abhishek Mohanty, date: 10/20/2019, view: 260, Q&A: 0
    [Abstract] Mammalian skeletal muscle is a metabolically active tissue that is made up of different types of muscle fibers. These myofibers are made up of important contractile proteins that provide force during contraction of the muscle like actin and myosin. Murine myofibers have been classified into 4 types: Type I, Type IIa, Type IIb and Type IIX. Each ...
    3D Organoid Formation from the Murine Salivary Gland Cell Line SIMS
    Authors:  Harleen K. Athwal and Isabelle M. A. Lombaert, date: 10/05/2019, view: 360, Q&A: 0
    [Abstract] Salivary glands consist of multiple phenotypically and functionally unique cell populations, such as the acinar, ductal, and myoepithelial cells that help produce, modify, and secrete saliva (Lombaert et al., 2011). Identification of mechanisms and factors that regulate these populations has been of key interest, as salivary gland-related ...
    Reconstituting Breast Tissue with Organotypic Three-dimensional Co-culture of Epithelial and Stromal Cells in Discontinuous Extracellular Matrices
    Authors:  Gang Ren, Vandana Sharma, Joshua Letson, Yashna Walia, Veani Fernando and Saori Furuta, date: 10/05/2019, view: 432, Q&A: 0
    [Abstract] Co-culture systems utilizing reconstituted or synthetic extracellular matrix (ECM) and micropatterning techniques have enabled the reconstruction of surface epithelial tissues. This technique has been utilized in the regeneration, disease modeling and drug screening of the surface epithelia, such as the skin and esophagus. On the other hand, the ...
    A Widely Applicable Urea-based Fluorescent/Colorimetric mRNA in situ Hybridization Protocol
    Author:  Chiara Sinigaglia, date: 09/05/2019, view: 831, Q&A: 0
    [Abstract] In situ hybridization methods are routinely employed to detect nucleic acid sequences, allowing to localize gene expression or to study chromosomal organization in their native context. These methods rely on the pairwise binding of a labeled probe to the target endogenous nucleic acid sequence–the hybridization step, followed by detection ...
    Estrous Cycle Monitoring in Mice with Rapid Data Visualization and Analysis
    Authors:  Leanna K. Pantier, Jiang Li and Catherine A. Christian, date: 09/05/2019, view: 714, Q&A: 0
    [Abstract] The estrous cycle provides a readout of reproductive health in female laboratory rodents, and estrous cycle stage can be an important physiological variable. Accurate assessment of estrous cycle stage is also important in producing timed pregnancies for developmental studies. Here, we provide a protocol for evaluation of estrous cycle stage ...
    Application of a Modified Smart-seq2 Sample Preparation Protocol for Rare Cell Full-length Single-cell mRNA Sequencing to Mouse Oocytes
    Authors:  Rebecca S. Treger, Scott D. Pope, Xiaojun Xing and Akiko Iwasaki, date: 08/20/2019, view: 609, Q&A: 0
    [Abstract] Endogenous retroviruses (ERV) are transposable retroelements that form ~10% of the murine genome and whose family members are differentially expressed throughout embryogenesis. However, precise regulation of ERV in germ cells remains unclear. To investigate ERV expression in oocytes, we adapted a single-cell mRNA-sequencing library preparation ...
    Ex vivo Drosophila Wing Imaginal Disc Culture and Furin Inhibitor Assay
    Authors:  Alex Sohr, Lijuan Du and Sougata Roy, date: 08/20/2019, view: 477, Q&A: 0
    [Abstract] Furin is an evolutionarily conserved proprotein convertase (PC) family enzyme with a broad range of substrates that are essential for developmental, homeostatic, and disease pathways. Classical genetic approaches and in vitro biochemical or cell biological assays identified that precursor forms of most growth factor family proteins are ...



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