Cell Biology

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    Protocols in Current Issue
    A Step-by-step Protocol for Obtaining Mature Microglia from Mice
    Authors:  Min-Jung Yoo and Min-Soo Kwon, date: 08/05/2022, view: 246, Q&A: 0
    [Abstract]

    In mice, microglial precursors in the yolk sac migrate to the brain parenchyma through the head neuroepithelial layer between embryonic days 8.5 (E8.5)–E16.5 and acquire their unique identity with a ramified form. Based on the microglial developmental process, we dissected the neuroepithelial layer (NEL) of E13.5 mice, which is composed

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    Protocol for Isolation of Cardiomyocyte from Adult Mouse and Rat
    Authors:  Huiliang Zhang and Peter S. Rabinovitch, date: 05/20/2022, view: 1086, Q&A: 0
    [Abstract]

    The isolation of intact single adult cardiomyocytes from model animals, mouse and rat, is an essential tool for cardiac molecular and cellular research. While several methods are reported for adult mouse cardiomyocyte isolation, the viability and yield of the isolated cells have been variable. Here, we describe step-by-step protocols for high

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    Isolation and Culture of Cranial Neural Crest Cells from the First Branchial Arch of Mice
    Authors:  Hiroki Ueharu, Jingwen Yang, Yoshihiro Komatsu and Yuji Mishina, date: 04/05/2022, view: 774, Q&A: 1
    [Abstract]

    Craniofacial anomalies (CFA) are a diverse group of deformities, which affect the growth of the head and face. Dysregulation of cranial neural crest cell (NCC) migration, proliferation, differentiation, and/or cell fate specification have been reported to contribute to CFA. Understanding of the mechanisms through which cranial NCCs contribute

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    Differentiation of Human Induced Pluripotent Stem Cell into Macrophages
    Authors:  Harriet Douthwaite, Aitor Bermejo Arteagabeitia and Subhankar Mukhopadhyay, date: 03/20/2022, view: 1242, Q&A: 0
    [Abstract]

    As a model to interrogate human macrophage biology, macrophages differentiated from human induced pluripotent stem cells (hiPSCs) transcend other existing models by circumventing the variability seen in human monocyte-derived macrophages, whilst epitomizing macrophage phenotypic and functional characteristics over those offered by macrophage-like

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    Efficient Method to Differentiate Mouse Embryonic Stem Cells into Macrophages in vitro
    Authors:  Qimin Hai, Juying Han, Sophia Wells and Jonathan D. Smith, date: 02/05/2022, view: 1597, Q&A: 0
    [Abstract]

    Macrophages are key cells in the innate immune system and play a role in a variety of diseases. However, macrophages are terminally differentiated and difficult to manipulate genetically via transfection or through CRISPR-Cas9 gene editing. To overcome this limitation, we provide a simplified protocol for the generation of mouse embryonic stem

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    A Method to Induce Brown/Beige Adipocyte Differentiation from Murine Preadipocytes
    Authors:  Andréa Livia Rocha, Beatriz Alves Guerra, Jeremie Boucher and Marcelo A. Mori, date: 12/20/2021, view: 1619, Q&A: 0
    [Abstract]

    Adipocytes exhibit different morphological and functional characteristics, depending on their anatomical location, developmental origin, and stimulus. While white adipocytes tend to accumulate energy as triglycerides, brown and beige adipocytes tend to direct carbon sources to fuel thermogenesis. White and beige adipocytes originate from common

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    High-throughput 3D Spheroid Formation and Effective Cardiomyocyte Differentiation from Human iPS Cells Using the Microfabric Vessels EZSPHERETM
    Authors:  Tatsuaki Miwa, Alimjan Idiris and Hiromichi Kumagai, date: 11/05/2021, view: 1402, Q&A: 0
    [Abstract]

    High-throughput 3D spheroid formation from human induced pluripotent stem cells (hiPSCs) can be easily performed using the unique microfabric vessels EZSPHERE, resulting in effective and large scale generation of differentiated cells such as cardiomyocytes or neurons. Such hiPSC-derived cardiomyocytes (hiPSC-CMs) or neurons are very useful in the

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    Mouse Periosteal Cell Culture, in vitro Differentiation, and in vivo Transplantation in Tibial Fractures
    [Abstract]

    The periosteum covering the outer surface of bone contains skeletal stem/progenitor cells that can efficiently form cartilage and bone during bone repair. Several methods have been described to isolate periosteal cells based on bone scraping and/or enzymatic digestion. Here, we describe an explant culture method to isolate periosteum-derived

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    Mechanical Fractionation of Cultured Neuronal Cells into Cell Body and Neurite Fractions
    Authors:  Ankita Arora, Raeann Goering, Hei-Yong G. Lo and J. Matthew Taliaferro, date: 06/05/2021, view: 3003, Q&A: 0
    [Abstract]

    Many cells contain spatially defined subcellular regions that perform specialized tasks enabled by localized proteins. The subcellular distribution of these localized proteins is often facilitated by the subcellular localization of the RNA molecules that encode them. A key question in the study of this process of RNA localization is the

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    A Simple and Robust Protocol for in vitro Differentiation of Mouse Non-pathogenic T Helper 17 Cells from CD4+ T Cells
    Authors:  Siwen Kang, Ruohan Wu and Ruoning Wang, date: 05/20/2021, view: 3230, Q&A: 0
    [Abstract]

    Functional and mechanistic studies of CD4+ T cell lineages rely on robust methods of in vitro T cell polarization. Here, we report an optimized protocol for in vitro differentiation of a mouse non-pathogenic T helper 17 (TH17) cell lineage. Most of the previously established protocols require irradiated splenocytes as artificial antigen presenting

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