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    Protocols in Current Issue
    3D Culture Protocol for Testing Gene Knockdown Efficiency and Cell Line Derivation
    Authors:  Jan Strnadel, Sang Myung Woo, Sunkyu Choi, Huawei Wang, Marian Grendar and Ken Fujimura, date: 06/05/2018, view: 155, Q&A: 0
    Traditional 2D cell cultures with cells grown as monolayers on solid surface still represent the standard method in cancer research for drug testing. Cells grown in 2D cultures, however, lack relevant cell-matrix and cell-cell interactions and ...
    Hyaluronan Isolation from Mouse Mammary Gland
    Authors:  Cornelia Tolg, Mary Cowman and Eva A. Turley, date: 06/05/2018, view: 146, Q&A: 0
    The glycosaminoglycan hyaluronan (HA) is a key component of the extracellular matrix. The molecular weight of HA is heterogeneous and can reach from several million to several hundred daltons. The effect of HA on cell behavior is size dependent; ...
    3D Culture Protocol for Testing Gene Knockdown Efficiency and Cell Line Derivation
    Authors:  Jan Strnadel, Sang Myung Woo, Sunkyu Choi, Huawei Wang, Marian Grendar and Ken Fujimura, date: 06/05/2018, view: 155, Q&A: 0
    [Abstract] Traditional 2D cell cultures with cells grown as monolayers on solid surface still represent the standard method in cancer research for drug testing. Cells grown in 2D cultures, however, lack relevant cell-matrix and cell-cell interactions and ignore the true three-dimensional anatomy of solid tumors. Cells cultured in 2D can also undergo ...
    Hyaluronan Isolation from Mouse Mammary Gland
    Authors:  Cornelia Tolg, Mary Cowman and Eva A. Turley, date: 06/05/2018, view: 146, Q&A: 0
    [Abstract] The glycosaminoglycan hyaluronan (HA) is a key component of the extracellular matrix. The molecular weight of HA is heterogeneous and can reach from several million to several hundred daltons. The effect of HA on cell behavior is size dependent; fragmented HA acts as a danger signal, stimulates cell migration and proliferation and is ...
    Measurement of Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR) in Culture Cells for Assessment of the Energy Metabolism
    Authors:  Birte Plitzko and Sandra Loesgen, date: 05/20/2018, view: 695, Q&A: 0
    [Abstract] Mammalian cells generate ATP by mitochondrial (oxidative phosphorylation) and non-mitochondrial (glycolysis) metabolism. Cancer cells are known to reprogram their metabolism using different strategies to meet energetic and anabolic needs (Koppenol et al., 2011; Zheng, 2012). Additionally, each cancer tissue has its own individual ...
    An Image-based Assay for High-throughput Analysis of Cell Proliferation and Cell Death of Adherent Cells
    Authors:  Paula Szalai and Nikolai Engedal, date: 05/05/2018, view: 502, Q&A: 0
    [Abstract] In this protocol, we describe a method to monitor cell proliferation and death by live-cell imaging of propidium iodide (PI)-stained adherent mammalian cells. PI is widely used to assess cell death. However, it is usually used in end-point assays. Recently, we implemented the use of PI for real-time cell death assessment by automated imaging. ...
    Generation of Luciferase-expressing Tumor Cell Lines
    Authors:  Todd V. Brennan, Liwen Lin, Xiaopei Huang and Yiping Yang, date: 04/20/2018, view: 696, Q&A: 1
    [Abstract] Murine tumor models have been critical to advances in our knowledge of tumor physiology and for the development of effective tumor therapies. Essential to these studies is the ability to both track tumor development and quantify tumor burden in vivo. For this purpose, the introduction of genes that confer tumors with bioluminescent ...
    3D Co-culture System of Tumor-associated Macrophages and Ovarian Cancer Cells
    Authors:  Lingli Long, Mingzhu Yin and Wang Min, date: 04/20/2018, view: 652, Q&A: 0
    [Abstract] Ovarian cancer is fairly unique in that ovarian carcinoma cells can detach and spread directly through peritoneal cavity. It has been unclear, however, how detached cancer cells survive in the peritoneum and form spheroid structure. We have recently reported that there is a strong correlation between Tumor-associated macrophages (TAMs)-associated ...
    FACS-based Glucose Uptake Assay of Mouse Embryonic Fibroblasts and Breast Cancer Cells Using 2-NBDG Probe
    Authors:  Shengli Dong and Suresh K Alahari, date: 04/20/2018, view: 576, Q&A: 0
    [Abstract] This is a flow cytometry-based protocol to measure glucose uptake of mouse embryonic fibroblasts (MEFs) and breast cancer cells in vitro. The method is a slightly modified and updated version as previously described (Dong et al., 2017). Briefly, the target cells are incubated with the fluorescently tagged ...
    Centromere Chromosome Orientation Fluorescent in situ Hybridization (Cen-CO-FISH) Detects Sister Chromatid Exchange at the Centromere in Human Cells
    Author:  Simona Giunta, date: 04/05/2018, view: 770, Q&A: 0
    [Abstract] Human centromeres are composed of large tandem arrays of repetitive alpha satellite DNA, which are often sites of aberrant rearrangement in cancers (Mitelman et al., 1997; Padilla-Nash et al., 2001). To date, annotation of the human centromere repetitive sequences remains incomplete, greatly hindering in-depth functional studies ...
    Primary Cultures from Human GH-secreting or Clinically Non-functioning Pituitary Adenomas
    Authors:  Roberto Würth, Alessandra Pattarozzi, Federica Barbieri and Tullio Florio, date: 04/05/2018, view: 424, Q&A: 0
    [Abstract] Pituitary adenomas are among the more frequent intracranial tumors usually treated with both surgical and pharmacological–based on somatostatin and dopamine agonists–approaches. Although mostly benign tumors, the occurrence of invasive behaviors is often detected resulting in poorer prognosis. The use of primary cultures from human pituitary ...
    Detection and Analysis of Circular RNAs by RT-PCR
    Authors:  Amaresh C Panda and Myriam Gorospe, date: 03/20/2018, view: 1676, Q&A: 2
    [Abstract] Gene expression in eukaryotic cells is tightly regulated at the transcriptional and posttranscriptional levels. Posttranscriptional processes, including pre-mRNA splicing, mRNA export, mRNA turnover, and mRNA translation, are controlled by RNA-binding proteins (RBPs) and noncoding (nc)RNAs. The vast family of ncRNAs comprises diverse regulatory ...