Cell Biology

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    Protocols in Current Issue
    Isolation and in vitro Culture of Mouse Oocytes
    Authors:  Jessica Greaney, Goutham Narayanan Subramanian, Yunan Ye and Hayden Homer, date: 08/05/2021, view: 23, Q&A: 0
    [Abstract]

    Females are endowed at birth with a fixed reserve of oocytes, which declines both in quantity and quality with advancing age. Understanding the molecular mechanisms regulating oocyte quality is crucial for improving the chances of pregnancy success in fertility clinics. In vitro culture systems enable researchers to analyse important molecular and

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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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    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: 810, 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: 1254, 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: 949, 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: 1152, 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: 1574, 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: 1442, 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: 1843, 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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