Mammalia

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    Protocols in Current Issue
    A Method to Efficiently Cryopreserve Mammalian Cells on Paper Platforms
    Authors:  Muhammedin Deliorman, Pavithra Sukumar, Roaa Alnemari and Mohammad A. Qasaimeh, date: 09/20/2020, view: 291, Q&A: 0
    [Abstract] This protocol describes a simple method to cryopreserve mammalian cells within filter papers as an alternative to conventional slow-freezing approach. The method involves treating paper fibers with fibronectin, using low concentrations of the cryoprotectant dimethyl sulfoxide (DMSO), and slow freezing cells to -80 °C at a 1 °C min-1 ...
    Measuring Cell Growth and Junction Development in Epithelial Cells Using Electric Cell-Substrate Impedance Sensing (ECIS)
    Authors:  Shaista Anwer and Katalin Szaszi, date: 08/20/2020, view: 987, Q&A: 0
    [Abstract] Electric Cell-substrate Impedance Sensing (ECIS) is an automated method that can be used to quantify processes such as cell attachment, growth, migration and barrier functions (i.e., the properties of tight junctions). The method provides simultaneous information on cell number and tight junction function by detecting electric parameters ...
    Isolation of Tumor Cells Based on Their Distance from Blood Vessels
    Authors:  Saran Kumar, Husni Sharife, Tirzah Kreisel, Libat Bar-Lev, Myriam Grunewald and Eli Keshet, date: 05/20/2020, view: 1376, Q&A: 0
    [Abstract] Differential exposure of tumor cells to microenvironmental cues greatly impacts cell phenotypes, raising a need for position based sorting of tumor cells amenable to multiple OMICs and functional analyses. One such key determinant of tumor heterogeneity in solid tumors is its vasculature. Proximity to blood vessels (BVs) profoundly affects tumor ...
    Rapid Generation of Human Neuronal Cell Models Enabling Inducible Expression of Proteins-of-interest for Functional Studies
    [Abstract] CRISPR-Cas9 technology has transformed the ability to edit genomic sequences and control gene expression with unprecedented ease and scale. However, precise genomic insertions of coding sequences using this technology remain time-consuming and inefficient because they require introducing adjacent single-strand cuts through Cas9 nickase action and ...
    Model of Chemotherapy-associated Mucositis and Oral Opportunistic Infections
    Authors:  Takanori Sobue, Martinna Bertolini, Angela Thompson and Anna Dongari-Bagtzoglou, date: 11/05/2019, view: 1495, Q&A: 0
    [Abstract] Oral mucositis is a common complication of cancer chemotherapy treatment. Because of the lack of relevant oral mucositis experimental models, it is not clear how chemotherapeutic agents injure the oral mucosa and if commensal microorganisms accelerate tissue damage. We developed an organotypic oral mucosa model that mimics cellular responses ...
    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: 1796, 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 Novel Protocol to Generate Decellularized Bovine Spinal Cord Extracellular Matrix-based Scaffolds (3D-dCBS)
    Authors:  Yavuz E. Arslan, Burcu Efe and Tugba Sezgin Arslan, date: 10/05/2019, view: 1662, Q&A: 0
    [Abstract] Extracellular matrix (ECM)-based tissue engineering scaffolds have an essential role in promoting tissue regeneration. Nerve tissue engineering aims at facilitating the repair of permanent damage to the peripheral and central nervous systems, which are difficult to heal. For this purpose, a variety of biomaterials are being developed consisting of ...
    One-step Derivation of Functional Mesenchymal Stem Cells from Human Pluripotent Stem Cells
    Authors:  Yan Zhou, JinQi Liao, ChongZhou Fang, CuiPing Mo, GuangQian Zhou and Yonglun Luo, date: 11/20/2018, view: 3949, Q&A: 0
    [Abstract] Mesenchymal stem cells (MSCs) are invaluable cell sources for understanding stem cell biology and potential application in tissue engineering and regenerative medicine. The current issues of MSCs that demand to be further addressed are limited donors, tissue sources and limited capacity of ex vivo expansion. Here, we describe a simple and ...
    Generation of Human Mesenchymal Stem Cell 3D Spheroids Using Low-binding Plates
    [Abstract] The 3D culture of human mesenchymal stem cells (hMSCs) represents a more physiological environment than classical 2D culture and has been used to enhance the MSC secretome or extend cell survival after transplantation. Here we describe a simple and affordable method to generate 3D spheroids of hMSCs by seeding them at high density in a low-binding ...
    Generating Loss-of-function iPSC Lines with Combined CRISPR Indel Formation and Reprogramming from Human Fibroblasts
    Authors:  Andrew M. Tidball, Preethi Swaminathan, Louis T. Dang and Jack M. Parent, date: 04/05/2018, view: 6960, Q&A: 1
    [Abstract] For both disease and basic science research, loss-of-function (LOF) mutations are vitally important. Herein, we provide a simple stream-lined protocol for generating LOF iPSC lines that circumvents the technical challenges of traditional gene-editing and cloning of established iPSC lines by combining the introduction of the CRISPR vector ...



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