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Cancer Biology

Lentiviral shRNA Screen to Identify Epithelial Integrity Regulating Genes in MCF10A 3D Culture

Featured protocol,  Authors: Elsa Marques
Elsa Marques Affiliation: Cancer Cell Circuitry Laboratory, Research Programs Unit/Translational Cancer Biology, University of Helsinki, Helsinki, Finland
Bio-protocol author page: a3834
 and Juha Klefström
Juha KlefströmAffiliation: Cancer Cell Circuitry Laboratory, Research Programs Unit/Translational Cancer Biology, University of Helsinki, Helsinki, Finland
For correspondence: juha.klefstrom@helsinki.fi
Bio-protocol author page: a3835
date: 12/5/2016, 71 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.2050.

Brief version appeared in Oncogene, Mar 2016
MCF10A 3D culture system provides a reductionist model of glandular mammary epithelium which is widely used to study development of glandular architecture, the role of cell polarity and epithelial integrity in control of epithelial cell functions, and mechanisms of breast cancer. Here we describe how to use shRNA screening approach to identify critical cell pathways that couple epithelial structure to individual cell based responses such as cell cycle exit and apoptosis. These studies will help to interrogate genetic changes critical for early breast tumorigenesis. The protocol describes a library of lentiviral shRNA constructs designed to target epithelial integrity and a highly efficient method for lentiviral transduction of suspension MCF10A cultures. Furthermore, protocols are provided for setting up MCF10A 3D cultures in Matrigel for morphometric and cellular response studies via structured illumination and confocal microscopy analysis of immunostained 3D structures.

DNA Damage Induction by Laser Microirradiation

Featured protocol,  Authors: Marianna Tampere
Marianna TampereAffiliation: Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden
Bio-protocol author page: a3807
 and Oliver Mortusewicz
Oliver MortusewiczAffiliation: Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden
For correspondence: oliver.mortusewicz@scilifelab.se
Bio-protocol author page: a3808
date: 12/5/2016, 46 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.2039.

Brief version appeared in Oncogene, Feb 2016
Genome instability can lead to cell death, senescence and cancerous transformation. Specific repair pathways have evolved to prevent accumulation of DNA lesions. Studying these highly dynamic and specific repair pathways requires precise spatial and temporal resolution, which can be achieved through a combination of laser microirradiaiton and live cell microscopy. DNA lesions are introduced at pre-determined sub-nuclear sites and repair can be analyzed in real time in living cells when using fluorescently tagged repair proteins (Mortusewicz et al., 2008). Alternatively, laser microirradiation can be combined with immunofluorescence analysis to study recruitment of endogenous proteins to laser-induced DNA damage tracks that can be visualized by positive controls like, e.g., γH2AX that mark sites of DNA breaks.

Establishment of Patient-Derived Xenografts in Mice

Featured protocol,  Authors: Dongkyoo Park
Dongkyoo ParkAffiliation: Division of Cancer Biology, Department of Radiation Oncology, Emory University School of Medicine and Winship Cancer Institute of Emory University, Atlanta, GA, USA
Bio-protocol author page: a3712
Dongsheng Wang
Dongsheng WangAffiliation: Division of Cancer Biology, Department of Radiation Oncology, Emory University School of Medicine and Winship Cancer Institute of Emory University, Atlanta, GA, USA
Bio-protocol author page: a3713
Guo Chen
Guo ChenAffiliation: Division of Cancer Biology, Department of Radiation Oncology, Emory University School of Medicine and Winship Cancer Institute of Emory University, Atlanta, GA, USA
Bio-protocol author page: a3714
 and Xingming Deng
Xingming DengAffiliation: Division of Cancer Biology, Department of Radiation Oncology, Emory University School of Medicine and Winship Cancer Institute of Emory University, Atlanta, GA, USA
For correspondence: xdeng4@emory.edu
Bio-protocol author page: a3715
date: 11/20/2016, 189 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.2008.

Brief version appeared in Cancer Cell, Jun 2015
Patient-derived xenograft (PDX) models for cancer research have recently attracted considerable attention in both the academy and industry (Hidalgo et al., 2014; Wilding and Bodmer, 2014). PDX models have been developed from different tumor types including lung cancer to improve the drug development process. These models are used for pre-clinical drug evaluation and can be used for the predictive results of clinical outcomes because they conserve original tumor characteristics such as heterogeneity, complexity and molecular diversity (Kopetz et al., 2012). Additionally, PDX model provides the potential tool for the personalized drug therapy. In this protocol, we present methods for the establishment of PDX in mice using primary tumor tissues from patients with small cell lung cancer (SCLC).

Total Histone Acid Extraction of Colon Cancer HCT116 Cells

Featured protocol,  Authors: Lin-Lin Cao
Lin-Lin CaoAffiliation: Department of Clinical Laboratory, Peking University People’s Hospital, Beijing, China
Bio-protocol author page: a3751
 and Wei-Guo Zhu
Wei-Guo ZhuAffiliation 1: Department of Biochemistry and Molecular Biology, School of Medicine, Shenzhen University, Shenzhen, China
Affiliation 2: Peking University-Tsinghua University Joint Center for Life Sciences, Beijing, China
For correspondence: zhuweiguo@szu.edu.cn
Bio-protocol author page: a3752
date: 11/20/2016, 117 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.2023.

Brief version appeared in Oncogene, Jan 2016
Histone acid extraction assay is a popular method to determine histone modification levels in mammalian cells. It includes three steps: first, histones are released from chromatin by sulfuric acid; trichloroacetate (TCA) is then added to precipitate histones; and finally, histones are dissolved in double-distilled H2O (ddH2O). Here we present a detailed histone acid extraction assay in our laboratory using a colon cancer cell line, HCT116, as a model.

Determining the Influence of Small Molecules on Hypoxic Prostate Cancer Cell (DU-145) Viability Using Automated Cell Counting and a Cell Harvesting Protocol

Featured protocol,  Authors: John P Phelan
John P PhelanAffiliation: Biomerit Research Centre, School of Microbiology, University College Cork, Cork, Ireland
Bio-protocol author page: a3733
F Jerry Reen
F Jerry ReenAffiliation: Biomerit Research Centre, School of Microbiology, University College Cork, Cork, Ireland
Bio-protocol author page: a3775
 and Fergal O’Gara
Fergal O’GaraAffiliation 1: Biomerit Research Centre, School of Microbiology, University College Cork, Cork, Ireland
Affiliation 2: School of Biomedical Sciences, Curtin University, Perth, Australia
For correspondence: f.ogara@ucc.ie
Bio-protocol author page: a3734
date: 11/20/2016, 109 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.2017.

Brief version appeared in BMC Cancer, Jul 2016
Cell viability assays are an essential aspect of most cancer studies, however they usually require a considerable labor and time input. Here, instead of using the conventional microscopy and hemocytometer cell counting approach, we developed a cell harvesting protocol and combined it with the automated Countess Automated Cell Counter to generate cell viability data. We investigated the effects of dihydroxylated bile acids on the cell viability of prostate cancer cells grown under hypoxic conditions. We observed that for all conditions, cell viability was relatively unchanged, suggesting these molecules had little or no impact on cell viability. The combination of the automated approach and the cell harvesting protocol means this assay is i) easy to perform, ii) extremely reproducible and iii) it complements more conventional cancer assay data such as invasion, migration and adhesion.

Lentiviral shRNA Screen to Identify Epithelial Integrity Regulating Genes in MCF10A 3D Culture

Authors: Elsa Marques
Elsa Marques Affiliation: Cancer Cell Circuitry Laboratory, Research Programs Unit/Translational Cancer Biology, University of Helsinki, Helsinki, Finland
Bio-protocol author page: a3834
 and Juha Klefström
Juha KlefströmAffiliation: Cancer Cell Circuitry Laboratory, Research Programs Unit/Translational Cancer Biology, University of Helsinki, Helsinki, Finland
For correspondence: juha.klefstrom@helsinki.fi
Bio-protocol author page: a3835
date: 12/5/2016, 71 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.2050.

[Abstract] MCF10A 3D culture system provides a reductionist model of glandular mammary epithelium which is widely used to study development of glandular architecture, the role of cell polarity and epithelial integrity in control of epithelial cell functions, and mechanisms of breast cancer. Here we describe how to use shRNA screening approach to identify critical ...

DNA Damage Induction by Laser Microirradiation

Authors: Marianna Tampere
Marianna TampereAffiliation: Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden
Bio-protocol author page: a3807
 and Oliver Mortusewicz
Oliver MortusewiczAffiliation: Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden
For correspondence: oliver.mortusewicz@scilifelab.se
Bio-protocol author page: a3808
date: 12/5/2016, 46 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.2039.

[Abstract] Genome instability can lead to cell death, senescence and cancerous transformation. Specific repair pathways have evolved to prevent accumulation of DNA lesions. Studying these highly dynamic and specific repair pathways requires precise spatial and temporal resolution, which can be achieved through a combination of laser microirradiaiton and live ...

Establishment of Patient-Derived Xenografts in Mice

Authors: Dongkyoo Park
Dongkyoo ParkAffiliation: Division of Cancer Biology, Department of Radiation Oncology, Emory University School of Medicine and Winship Cancer Institute of Emory University, Atlanta, GA, USA
Bio-protocol author page: a3712
Dongsheng Wang
Dongsheng WangAffiliation: Division of Cancer Biology, Department of Radiation Oncology, Emory University School of Medicine and Winship Cancer Institute of Emory University, Atlanta, GA, USA
Bio-protocol author page: a3713
Guo Chen
Guo ChenAffiliation: Division of Cancer Biology, Department of Radiation Oncology, Emory University School of Medicine and Winship Cancer Institute of Emory University, Atlanta, GA, USA
Bio-protocol author page: a3714
 and Xingming Deng
Xingming DengAffiliation: Division of Cancer Biology, Department of Radiation Oncology, Emory University School of Medicine and Winship Cancer Institute of Emory University, Atlanta, GA, USA
For correspondence: xdeng4@emory.edu
Bio-protocol author page: a3715
date: 11/20/2016, 189 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.2008.

[Abstract] Patient-derived xenograft (PDX) models for cancer research have recently attracted considerable attention in both the academy and industry (Hidalgo et al., 2014; Wilding and Bodmer, 2014). PDX models have been developed from different tumor types including lung cancer to improve the drug development process. These models are used for pre-clinical drug ...

Total Histone Acid Extraction of Colon Cancer HCT116 Cells

Authors: Lin-Lin Cao
Lin-Lin CaoAffiliation: Department of Clinical Laboratory, Peking University People’s Hospital, Beijing, China
Bio-protocol author page: a3751
 and Wei-Guo Zhu
Wei-Guo ZhuAffiliation 1: Department of Biochemistry and Molecular Biology, School of Medicine, Shenzhen University, Shenzhen, China
Affiliation 2: Peking University-Tsinghua University Joint Center for Life Sciences, Beijing, China
For correspondence: zhuweiguo@szu.edu.cn
Bio-protocol author page: a3752
date: 11/20/2016, 117 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.2023.

[Abstract] Histone acid extraction assay is a popular method to determine histone modification levels in mammalian cells. It includes three steps: first, histones are released from chromatin by sulfuric acid; trichloroacetate (TCA) is then added to precipitate histones; and finally, histones are dissolved in double-distilled H2O (ddH2O). Here we present a detailed ...

Determining the Influence of Small Molecules on Hypoxic Prostate Cancer Cell (DU-145) Viability Using Automated Cell Counting and a Cell Harvesting Protocol

Authors: John P Phelan
John P PhelanAffiliation: Biomerit Research Centre, School of Microbiology, University College Cork, Cork, Ireland
Bio-protocol author page: a3733
F Jerry Reen
F Jerry ReenAffiliation: Biomerit Research Centre, School of Microbiology, University College Cork, Cork, Ireland
Bio-protocol author page: a3775
 and Fergal O’Gara
Fergal O’GaraAffiliation 1: Biomerit Research Centre, School of Microbiology, University College Cork, Cork, Ireland
Affiliation 2: School of Biomedical Sciences, Curtin University, Perth, Australia
For correspondence: f.ogara@ucc.ie
Bio-protocol author page: a3734
date: 11/20/2016, 109 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.2017.

[Abstract] Cell viability assays are an essential aspect of most cancer studies, however they usually require a considerable labor and time input. Here, instead of using the conventional microscopy and hemocytometer cell counting approach, we developed a cell harvesting protocol and combined it with the automated Countess Automated Cell Counter to generate cell ...

Fat Turnover Assay in Drosophila

Author: Subhash D. Katewa
Subhash D. KatewaAffiliation: Buck Institute for Research on Aging, Novato, CA, USA
For correspondence: skatewa@buckinstitute.org
Bio-protocol author page: a3681
date: 11/5/2016, 212 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.1996.

[Abstract] Like all animals, Drosophila shows robust fat (triglyceride) turnover, i.e., they synthesize, store and utilize triglyceride for their daily metabolic needs. The protocol describes a simple assay to measure this turnover of triglycerides in Drosophila.

[Background] Almost all animals store energy reserves in the form of glycogen and triglycerides. Many ...

Sulforhodamine B (SRB) Assay in Cell Culture to Investigate Cell Proliferation

Authors: Esteban A. Orellana
Esteban A. OrellanaAffiliation 1: Department of Biological Sciences, Bindley Bioscience Center, Purdue University, West Lafayette, USA
Affiliation 2: Purdue University Interdisciplinary Life Science Program (PULSe), Purdue University, West Lafayette, USA
Bio-protocol author page: a3649
 and Andrea L. Kasinski
Andrea L. KasinskiAffiliation: Department of Biological Sciences, Bindley Bioscience Center, Purdue University, West Lafayette, USA
For correspondence: akasinski@purdue.edu
Bio-protocol author page: a380
date: 11/5/2016, 216 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.1984.

[Abstract] The SRB assay has been used since its development in 1990 (Skehan et al., 1990) to inexpensively conduct various screening assays to investigate cytotoxicity in cell based studies (Vichai and Kirtikara, 2006). This method relies on the property of SRB, which binds stoichiometrically to proteins under mild acidic conditions and then can be extracted ...

Phagocytosis Assay to Measure Uptake of Necroptotic Cancer Cells by BMDCs

Authors: Tania Løve Aaes
Tania Løve AaesAffiliation 1: Molecular Signaling and Cell Death Unit, VIB Inflammation Research Center, Ghent, Belgium
Affiliation 2: Department of Biomedical Molecular Biology, Cancer Research Institute Ghent (CRIG), Ghent University, Ghent, Belgium
Bio-protocol author page: a3682
Dmitri V. Krysko
Dmitri V. KryskoAffiliation 1: Molecular Signaling and Cell Death Unit, VIB Inflammation Research Center, Ghent, Belgium
Affiliation 2: Department of Biomedical Molecular Biology, Cancer Research Institute Ghent (CRIG), Ghent University, Ghent, Belgium
Affiliation 3: Department of Basic Medical Sciences, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium
Bio-protocol author page: a3683
 and Peter Vandenabeele
Peter VandenabeeleAffiliation 1: Molecular Signaling and Cell Death Unit, VIB Inflammation Research Center, Ghent, Belgium
Affiliation 2: Department of Biomedical Molecular Biology, Cancer Research Institute Ghent (CRIG), Ghent University, Ghent, Belgium
Affiliation 3: Methusalem Program, Ghent University, Ghent, Belgium
For correspondence: peter.vandenabeele@irc.vib-ugent.be
Bio-protocol author page: a3684
date: 11/5/2016, 190 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.1997.

[Abstract] This protocol is a flow cytometry-based method to measure the phagocytosis efficiency of necroptotic target cells by bone marrow-derived dendritic cells (BMDCs) in vitro (Aaes et al., 2016). The method is a slightly modified and updated version of the protocols used in previously published papers (Krysko et al., 2006; Brouckaert et al., 2004). In brief, ...

Quantification of Tumor Material Uptake

Authors: Richard N. Hanna
Richard N. HannaAffiliation 1: Department of Respiratory, Inflammation and Autoimmune Diseases, MedImmune, LLC, Gaithersburg, USA
Affiliation 2: Division of Inflammation Biology, La Jolla Institute for Allergy and Immunology, La Jolla, USA
For correspondence: hannar@Medimmune.com
Bio-protocol author page: a3623
 and Catherine C. Hedrick
Catherine C. HedrickAffiliation: Division of Inflammation Biology, La Jolla Institute for Allergy and Immunology, La Jolla, USA
Bio-protocol author page: a3625
date: 10/20/2016, 284 views, 1 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.1974.

[Abstract] Extracellular tumor material including exosomes, microvesicles and apoptotic tumor debris may help cancers invade new organs. Enhancing the removal of extracellular tumor material by immune cells represents a novel immunotherapy approach for preventing cancer metastasis. This protocol quantifies the uptake and removal of extracellular tumor material ...

In vivo Imaging of Tumor and Immune Cell Interactions in the Lung

Authors: Richard N. Hanna
Richard N. HannaAffiliation 1: Department of Respiratory, Inflammation and Autoimmune Diseases, MedImmune, LLC, Gaithersburg, USA
Affiliation 2: Division of Inflammation Biology, La Jolla Institute for Allergy and Immunology, La Jolla, USA
For correspondence: hannar@Medimmune.com
Bio-protocol author page: a3623
Grzegorz Chodaczek
Grzegorz ChodaczekAffiliation: Microscopy Core, La Jolla Institute for Allergy and Immunology, La Jolla, USA
Bio-protocol author page: a3624
 and Catherine C. Hedrick
Catherine C. HedrickAffiliation: Division of Inflammation Biology, La Jolla Institute for Allergy and Immunology, La Jolla, USA
Bio-protocol author page: a3625
date: 10/20/2016, 351 views, 0 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.1973.

[Abstract] Immunotherapy has demonstrated great therapeutic potential by activating the immune system to fight cancer. However, little is known about the specific dynamics of interactions that occur between tumor and immune cells. In this protocol we describe a novel method to visualize the interaction of tumor and immune cells in the lung of live mice, which ...
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Scratch Wound Healing Assay

Author: Yanling Chen
Yanling ChenAffiliation: Department of Immunology, The Scripps Research Institute, La Jolla, USA
For correspondence: ylchen@scripps.edu
Bio-protocol author page: a27
date: 3/5/2012, 50813 views, 6 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.100.

[Abstract] The scratch wound healing assay has been widely adapted and modified to study the effects of a variety of experimental conditions, for instance, gene knockdown or chemical exposure, on mammalian cell migration and proliferation. In a typical scratch wound healing assay, a “wound gap” in a cell monolayer ...

Transwell Cell Migration Assay Using Human Breast Epithelial Cancer Cell

Author: Yanling Chen
Yanling ChenAffiliation: Department of Immunology, The Scripps Research Institute, La Jolla, USA
For correspondence: ylchen@scripps.edu
Bio-protocol author page: a27
date: 2/20/2012, 41083 views, 7 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.99.

[Abstract] Transwell migration assays have been widely used for studying the motility of different types of cells including metastatic cancer cells. The assay is also useful in screens for compounds that act as chemoattractants or inhibitors of chemotaxis for cells. The assay employs a permeable layer of support, ...

[Bio101] Cell Proliferation Assay by Flow Cytometry (BrdU and PI Staining)

Author: Hui Zhu
Hui ZhuAffiliation: Department of Genetics, Stanford University, Stanford, USA
For correspondence: huizhu@stanford.edu
Bio-protocol author page: a32
date: 4/5/2012, 36669 views, 2 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.198.

[Abstract] Cell Proliferation assays include an important set of fluorescence-based tests that can monitor cell health and cell division by evaluating DNA synthesis through thymidine incorporation. Bromodeoxyuridine (5-bromo-2'-deoxyuridine, BrdU) is a synthetic nucleoside that is an analogue of thymidine. BrdU ...

Clonogenic Assay

Author: Xiaodong Yang
Xiaodong YangAffiliation: Department of Neurology, University of California, San Francisco, USA
For correspondence: yangxiaodong1@yahoo.com
Bio-protocol author page: a43
date: 5/20/2012, 34055 views, 5 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.187.

[Abstract] Clonogenic assays serve as a useful tool to test whether a given cancer therapy can reduce the clonogenic survival of tumor cells. A colony is defined as a cluster of at least 50 cells that can often only be determined microscopically. A clonogenic assay is the method of choice to determine cell reproductive ...

In vitro Human Umbilical Vein Endothelial Cells (HUVEC) Tube-formation Assay

Authors: Josephine MY Ko
Josephine MY KoAffiliation: Clinical Oncology Department, The University of Hong Kong, Hong Kong , Hong Kong SAR
Bio-protocol author page: a100
 and Maria Li Lung
Maria Li LungAffiliation: Clinical Oncology Department, The Univerisity of Hong Kong, Hong Kong, Hong Kong SAR
For correspondence: mlilung@hku.hk
Bio-protocol author page: a101
date: 9/20/2012, 30116 views, 1 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.260.

[Abstract] Angiogenesis is involved not only in pathological conditions including cancer biology and non-neoplastic diseases, but also many biological processes including reproduction, development and repair. During angiogenesis, endothelial cells (ECs) undergo activation after binding of angiogenic factors to ...

Soft–Agar colony Formation Assay

Author: FengZhi Liu
FengZhi LiuAffiliation: School of Biomedical Sciences, Thomas Jefferson University, Philadelphia, USA
For correspondence: fengzhi6@yahoo.com
Bio-protocol author page: a51
date: 7/5/2012, 29540 views, 6 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.220.

[Abstract] Any anchorage–independent growth of tumor cells is estimated by a soft–agar colony formation assay. This protocol provides a general workflow for establishing a soft-agar colony formation assay....

[Bio101] Cell Adhesion Assay

Author: Yanling Chen
Yanling ChenAffiliation: Department of Immunology, The Scripps Research Institute, La Jolla, USA
For correspondence: ylchen@scripps.edu
Bio-protocol author page: a27
date: 3/5/2012, 25397 views, 1 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.98.

[Abstract] Cell adhesion, the binding of a cell to the extracellular matrix (ECM), other cells, or a specific surface, is essential for the growth and survival of the cell and also its communication with other cells. The process of cell adhesion involves a range of biological events such as three-dimensional re-organization ...

[Bio101] Subcutaneous Injection of Tumor Cells

Author: Jason Reuter date: 12/20/2011, 22290 views, 1 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.166.

[Abstract] Growth of cells in the subcutaneous space of immunocompromised mice is a common method for assaying tumorigenic potential in vivo. This technique is also used to assess the effects of therapeutic interventions on cancer cell lines....

In vitro Tumorsphere Formation Assays

Authors: Sara Johnson
Sara JohnsonAffiliation: Biological Sciences Department, University of South Carolina, Columbia, USA
Bio-protocol author page: a224
Hexin Chen
Hexin ChenAffiliation: Biological Sciences Department, University of South Carolina, Columbia, USA
For correspondence: hchen@biol.sc.edu
Bio-protocol author page: a225
 and Pang-Kuo Lo
Pang-Kuo LoAffiliation: Biological Sciences Department, University of South Carolina, Columbia, USA
Bio-protocol author page: a226
date: 2/5/2013, 21924 views, 3 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.325.

[Abstract] A tumorsphere is a solid, spherical formation developed from the proliferation of one cancer stem/progenitor cell. These tumorspheres (Figure 1a) are easily distinguishable from single or aggregated cells (Figure 1b) as the cells appear to become fused together and individual cells cannot be identified. ...

In vivo Matrigel Plug Angiogenesis Assay

Authors: Hong Lok Lung
Hong Lok LungAffiliation: Department of Clinical Oncology and Center for Cancer Research, The University of Hong Kong, Hong Kong , Hong Kong SAR
Bio-protocol author page: a102
 and Maria Li Lung
Maria Li LungAffiliation: Department of Clinical Oncology and Center for Cancer Research, The University of Hong Kong, Hong Kong , Hong Kong SAR
For correspondence: mlilung@hku.hk
Bio-protocol author page: a101
date: 9/20/2012, 21892 views, 2 Q&A, How to cite
DOI: https://doi.org/10.21769/BioProtoc.261.

[Abstract] The matrigel plug angiogenesis assay is a simple in vivo technique to detect the newly formed blood vessels in the transplanted gel plugs in nude mice. The matrigel matrix is derived from the engelbroth-holm-swarm (EHS) mouse sarcoma, and its composition is comparable to the basement membrane proteins. ...
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