Cell Biology

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    Protocols in Current Issue
    Preparation of Drosophila Larval Blood Cells for Single-cell RNA Sequencing
    Authors:  Sudhir Gopal Tattikota and Norbert Perrimon, date: 08/20/2021, view: 464, Q&A: 0
    [Abstract]

    Recent advances in single-cell RNA-sequencing (scRNA-seq) technologies provide unprecedented opportunities to identify new cell types and characterize cell states. One of the most important requirements for performing scRNA-seq is to obtain high-quality single cells in suspension. Recently, we used this approach to characterize Drosophila blood

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    Isolation and in vitro Culture of Mouse Oocytes
    Authors:  Jessica Greaney, Goutham Narayanan Subramanian, Yunan Ye and Hayden Homer, date: 08/05/2021, view: 850, 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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    Enrichment of Vascular Fragments from Mouse Embryonic Brains for Endothelial Cell Analysis
    Authors:  Nicolas Santander and Thomas D. Arnold, date: 06/20/2021, view: 1070, 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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    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: 1708, 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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    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: 1434, 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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    Isolation of GFP-expressing Malarial Hypnozoites by Flow Cytometry Cell Sorting
    Authors:  Annemarie Voorberg-van der Wel, Sam O. Hoffman and Clemens H. M. Kocken, date: 05/05/2021, view: 1553, Q&A: 0
    [Abstract]

    Hypnozoites are dormant liver-stage parasites unique to relapsing malarial species, including the important human pathogen Plasmodium vivax, and pose a barrier to the elimination of malaria. Little is known regarding the biology of these stages, largely due to their inaccessible location. Hypnozoites can be cultured in vitro but these cultures

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    Measurements of Root Colonized Bacteria Species
    Authors:  Vílchez Juan Ignacio, Yu Yang, Dian Yi and Huiming Zhang, date: 04/05/2021, view: 2122, Q&A: 0
    [Abstract]

    Root-associated bacteria are able to influence plant fitness and vigor. A key step in understanding the belowground plant-bacteria interactions is to quantify root colonization by the bacteria of interest. Probably, genetic engineering with fluorescence markers is the most powerful way to monitor bacterial strains in plant. However, this could

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    Strategy of Isolating ‘Primed’ Tumor Initiating Cells Based on Mitochondrial Transmembrane Potential
    Authors:  Brian Spurlock, Vidya Sagar Hanumanthu and Kasturi Mitra, date: 03/05/2021, view: 2629, Q&A: 0
    [Abstract]

    Various stem cells have been found to be dependent on mitochondrial energetics. The role of mitochondria in regulating the self-renewal of normal stem cells and stem-like tumor initiating cells (TICs) is increasingly being appreciated. We proposed that TIC populations have a sub population of cells that are “primed” by mitochondria for

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    Isolation of Extracellular Vesicles Derived from Mesenchymal Stromal Cells by Ultracentrifugation
    [Abstract] Extracellular vesicles (EVs) are a heterogeneous group of membranous vesicles that differ on their biogenesis and release pathways, such as exosomes, microvesicles and apoptotic bodies. They are involved in cell-to-cell communication delivering signal molecules (proteins, nucleic acids, lipids, etc.) that can regulate different ...
    Dissecting the Rat Mammary Gland: Isolation, Characterization, and Culture of Purified Mammary Epithelial Cells and Fibroblasts
    [Abstract]

    With the advent of CRISPR-Cas and the ability to easily modify the genome of diverse organisms, rat models are being increasingly developed to interrogate the genetic events underlying mammary development and tumorigenesis. Protocols for the isolation and characterization of mammary epithelial cell subpopulations have been thoroughly developed for

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