Cell Biology

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    Protocols in Current Issue
    Preparation of Human Chondrocytes for Profiling Using Cytometry by Time-of-flight (cyTOF)
    Authors:  Fiorella Carla Grandi and Nidhi Bhutani, date: 07/20/2021, view: 64, Q&A: 0
    [Abstract]

    Single-cell technologies have allowed high-resolution profiling of tissues and thus a deeper understanding of tissue homeostasis and disease heterogeneity. Understanding this heterogeneity can be especially important for tailoring treatments in a patient-specific manner. Here, we detail methods for preparing human cartilage tissue for profiling

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    OrganoPlate Micro-fluidic Microvessel Culture and Analysis
    Authors:  Abidemi Junaid and Thomas Hankemeier, date: 07/05/2021, view: 1171, Q&A: 0
    [Abstract]

    The endothelial cells from the microvasculature are key drivers and targets of inflammatory and thrombotic processes in microvascular diseases. The study of bioactive lipids in inflammatory processes has been largely based on two-dimensional endothelial cell cultures. Three-dimensional microvessels-on-a-chip provides an opportunity to monitor the

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    Production, quantification, and infection of Amazonian Phlebovirus (Bunyaviridae)
    [Abstract]

    Phlebotomine vectors, sand flies of the order Diptera, are known to transmit Leishmania parasites as well as RNA viruses (arboviruses) to humans. The arbovirus, Icoaraci Phlebovirus (BeAN 24262 - ICOV), used in this study was isolated from Nectomys rodents, a mammalian species that is the same natural sylvatic reservoir of Leishmania (Leishmania)

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    Enrichment of Vascular Fragments from Mouse Embryonic Brains for Endothelial Cell Analysis
    Authors:  Nicolas Santander and Thomas D. Arnold, date: 06/20/2021, view: 913, Q&A: 0
    [Abstract]

    Endothelial cells in the brain interact with other cell types, forming the blood-brain barrier. This barrier controls the movement of solutes into and out of the brain, regulating pathophysiological processes and drug delivery to the brain. Common isolation methods used to study these cells during embryonic development involve enzymatic treatment

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    Quantitation of Secretory Granule Size in Drosophila Larval Salivary Glands
    Authors:  Cheng-I J. Ma and Julie A. Brill, date: 06/05/2021, view: 1116, Q&A: 0
    [Abstract]

    Maturation of secretory granules is a crucial process that ensures the bioactivity of cargo proteins undergoing regulated secretion. In Drosophila melanogaster, the larval salivary glands produce secretory granules that are up to four-fold larger in cross-sectional area after maturation. Therefore, we developed a live imaging microscopy approach

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    Isolation of First-Trimester and Full-term Human Placental Hofbauer Cells
    Authors:  Anna Appios, Jake R. Thomas and Naomi McGovern, date: 06/05/2021, view: 1470, Q&A: 0
    [Abstract]

    The placenta is the crucial organ that regulates the health of both mother and fetus during pregnancy. The human placenta is composed of villous tree-like structures that embed into the maternal decidua. Within the stroma of the villi resides a population of fetally-derived macrophages, the Hofbauer cells (HBC). HBC are the only fetal immune cells

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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: 1396, 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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    Generation and Maintenance of Homogeneous Human Midbrain Organoids
    Authors:  Henrik Renner, Martha Grabos, Hans R. Schöler and Jan M. Bruder, date: 06/05/2021, view: 1771, Q&A: 0
    [Abstract]

    Three-dimensional cell cultures (“organoids”) promise to better recapitulate native tissue physiology than traditional 2D cultures and are becoming increasingly interesting for disease modeling and compound screening efforts. While a number of protocols for the generation of neural organoids have been published, most protocols require

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    FACS Enrichment of Total Interstitial Cells and Fibroblasts from Adult Mouse Ventricles
    Authors:  Nona Farbehi, Vaibhao Janbandhu, Robert E. Nordon and Richard P. Harvey, date: 05/20/2021, view: 1174, Q&A: 0
    [Abstract]

    Besides cardiomyocytes, the heart contains numerous interstitial cell types, including cardiac fibroblasts, endothelial cells, immune (myeloid and lymphoid) cells, and mural cells (pericytes and vascular smooth muscle cells), which play key roles in heart repair, regeneration, and disease. We recently published a comprehensive map of cardiac

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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: 1701, 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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