Welcome guest, Sign in

Home

Cancer Biology

In vivo Efficacy Studies in Cell Line and Patient-derived Xenograft Mouse Models

Featured protocol,  Authors: Elizabeth A. Tovar
Elizabeth A. TovarAffiliation: Center for Cancer and Cell Biology, Van Andel Research Institute, Grand Rapids, Michigan, USA
Bio-protocol author page: a3966
Curt J. Essenburg
Curt J. EssenburgAffiliation: Center for Cancer and Cell Biology, Van Andel Research Institute, Grand Rapids, Michigan, USA
Bio-protocol author page: a3967
 and Carrie Graveel
Carrie GraveelAffiliation: Center for Cancer and Cell Biology, Van Andel Research Institute, Grand Rapids, Michigan, USA
For correspondence: carrie.graveel@vai.org
Bio-protocol author page: a3968
date: 1/5/2017, 87 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2100.

Brief version appeared in Clin Cancer Res, Feb 2016
In vivo xenograft models derived from human cancer cells have been a gold standard for evaluating the genetic drivers of cancer and are valuable preclinical models for evaluating the efficacy of cancer therapeutics. Recently, patient-derived tumorgrafts from multiple tumor types have been developed and shown to more accurately recapitulate the molecular and histological heterogeneity of cancer. Here we detail the procedures for developing patient-derived xenograft models from breast cancer tissue, cell-based xenograft models, serial tumor transplantation, tumor measurement, and drug treatment.

Measuring Procaspase-8 and -10 Processing upon Apoptosis Induction

Featured protocol,  Authors: Sabine Pietkiewicz
Sabine PietkiewiczAffiliation: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Present address: Medical advisory service of social health insurance, Essen, Germany
Bio-protocol author page: a3934
Clara Wolfe
Clara WolfeAffiliation: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Bio-protocol author page: a3935
Jörn H. Buchbinder
Jörn H. BuchbinderAffiliation: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Bio-protocol author page: a3936
 and Inna N. Lavrik
Inna N. LavrikAffiliation 1: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Affiliation 2: Federal research center Institute of Cytology and Genetics, Novosibirsk, Russia
For correspondence: inna.lavrik@med.ovgu.de
Bio-protocol author page: a3937
date: 1/5/2017, 97 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2081.

Brief version appeared in Cell Death Differ, Apr 2016
Apoptosis or programmed cell death is important for multicellular organisms to keep cell homeostasis and for the clearance of mutated or infected cells. Apoptosis can be induced by intrinsic or extrinsic stimuli. The first event in extrinsic apoptosis is the formation of the Death-Inducing Signalling Complex (DISC), where the initiator caspases-8 and -10 are fully activated by several proteolytic cleavage steps and induce the caspase cascade leading to apoptotic cell death. Analysing the processing of procaspases-8 and -10 by Western blot is a commonly used method to study the induction of apoptosis by death receptor stimulation. To analyse procaspase-8 and -10 cleavage, cells are stimulated with a death ligand for different time intervals, lysed and subjected to Western blot analysis using anti-caspase-8 and anti-caspase-10 antibodies. This allows monitoring the caspase cleavage products and thereby induction of apoptosis.

Generation of Tumour-stroma Minispheroids for Drug Efficacy Testing

Featured protocol,  Authors: Mark Watters
Mark Watters Affiliation: Apoptosis Research Centre, School of Natural Sciences, National University of Ireland, Galway, Ireland
Bio-protocol author page: a3964
 and Eva Szegezdi
Eva SzegezdiAffiliation: Apoptosis Research Centre, School of Natural Sciences, National University of Ireland, Galway, Ireland
For correspondence: eva.szegezdi@nuigalway.ie
Bio-protocol author page: a3965
date: 1/5/2017, 88 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2091.

Brief version appeared in Oncogene, Mar 2016
The three-dimensional organisation of cells in a tissue and their interaction with adjacent cells and extracellular matrix is a key determinant of cellular responses, including how tumour cells respond to stress conditions or therapeutic drugs (Elliott and Yuan, 2011). In vivo, tumour cells are embedded in a stroma formed primarily by fibroblasts that produce an extracellular matrix and enwoven with blood vessels. The 3D mixed cell type spheroid model described here incorporates these key features of the tissue microenvironment that in vivo tumours exist in; namely the three-dimensional organisation, the most abundant stromal cell types (fibroblasts and endothelial cells), and extracellular matrix. This method combined with confocal microscopy can be a powerful tool to carry out drug sensitivity, angiogenesis and cell migration/invasion assays of different tumour types.

Relative Stiffness Measurements of Cell-embedded Hydrogels by Shear Rheology in vitro

Featured protocol,  Authors: Thomas R. Cox
Thomas R. CoxAffiliation: The Garvan Institute of Medical Research and The Kinghorn Cancer Centre, Cancer Division, St Vincent's Clinical School, Faculty of Medicine, University of New South Wales, Sydney, Australia
For correspondence: t.cox@garvan.org.au
Bio-protocol author page: a3986
 and Chris D. Madsen
Chris D. MadsenAffiliation: Department of Laboratory Medicine, Division of Translational Cancer Research, Lund University, Lund, Sweden
For correspondence: chris.madsen@med.lu.se
Bio-protocol author page: a3987
date: 1/5/2017, 110 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2101.

Brief version appeared in EMBO Rep, Oct 2015
Hydrogel systems composed of purified extracellular matrix (ECM) components (such as collagen, fibrin, Matrigel, and methylcellulose) are a mainstay of cell and molecular biology research. They are used extensively in many applications including tissue regeneration platforms, studying organ development, and pathological disease models such as cancer. Both the biochemical and biomechanical properties influence cellular and tissue compatibility, and these properties are altered in pathological disease progression (Cox and Erler, 2011; Bonnans et al., 2014). The use of cell-embedded hydrogels in disease models such as cancer, allow the interrogation of cell-induced changes in the biomechanics of the microenvironment (Madsen et al., 2015). Here we report a simple method to measure these cell-induced changes in vitro using a controlled strain rotational rheometer.

In vitro Histone H3 Cleavage Assay for Yeast and Chicken Liver H3 Protease

Featured protocol,  Authors: Sakshi Chauhan
Sakshi ChauhanAffiliation: Laboratory of Chromatin Biology, Department of Biological Sciences, Indian Institute of Science Education and Research, Bhopal, India
Bio-protocol author page: a3946
Gajendra Kumar Azad
Gajendra Kumar AzadAffiliation 1: Laboratory of Chromatin Biology, Department of Biological Sciences, Indian Institute of Science Education and Research, Bhopal, India
Affiliation 2: Department of Genetics, Institute of Life Sciences, Hebrew University of Jerusalem, Jerusalem, Israel
Bio-protocol author page: a3947
 and Raghuvir Singh Tomar
Raghuvir Singh TomarAffiliation: Laboratory of Chromatin Biology, Department of Biological Sciences, Indian Institute of Science Education and Research, Bhopal, India
For correspondence: rst@iiserb.ac.in
Bio-protocol author page: a3948
date: 1/5/2017, 112 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2085.

Brief version appeared in Yeast, Jun 2016
Histone proteins are subjected to a wide array of reversible and irreversible post-translational modifications (PTMs) (Bannister and Kouzarides, 2011; Azad and Tomar, 2014). The PTMs on histones are known to regulate chromatin structure and function. Histones are irreversibly modified by proteolytic clipping of their tail domains. The proteolytic clipping of histone tails is continuously attracting interest of researchers in the field of chromatin biology. We can recapitulate H3-clipping by performing in vitro H3 cleavage assay. Here, we are presenting the detailed protocol to perform in vitro H3 cleavage assay.

Affinity Pulldown of Biotinylated RNA for Detection of Protein-RNA Complexes

Featured protocol,  Authors: Amaresh C Panda
Amaresh C PandaAffiliation: Laboratory of Genetics and Genomics, National Institute on Aging, National Institutes of Health, 251 Bayview Blvd., Baltimore, United States
For correspondence: amaresh.panda@nih.gov
Bio-protocol author page: a3875
Jennifer L. Martindale
Jennifer L. Martindale Affiliation: Laboratory of Genetics and Genomics, National Institute on Aging, National Institutes of Health, 251 Bayview Blvd., Baltimore, United States
Bio-protocol author page: a3880
 and Myriam Gorospe
Myriam GorospeAffiliation: Laboratory of Genetics and Genomics, National Institute on Aging, National Institutes of Health, 251 Bayview Blvd., Baltimore, United States
Bio-protocol author page: a3881
date: 12/20/2016, 228 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2062.

Brief version appeared in Nucleic Acids Res, Mar 2016
RNA-binding proteins (RBPs) have recently emerged as crucial players in the regulation of gene expression. The interactions of RBPs with target mRNAs control the levels of gene products by altering different regulatory steps, including pre-mRNA splicing and maturation, nuclear mRNA export, and mRNA stability and translation (Glisovic et al., 2008). There are several methodologies available today to identify RNAs bound to specific RBPs; some detect only recombinant molecules in vitro, others detect recombinant and endogenous molecules, while others detect only endogenous molecules. Examples include systematic evolution of ligands by exponential enrichment (SELEX), biotinylated RNA pulldown assay, RNA immunoprecipitation (RIP) assay, electrophoretic mobility shift assay (EMSA), RNA footprinting analysis, and various UV crosslinking and immunoprecipitation (CLIP) methods such as CLIP, PAR-CLIP, and iCLIP (Popova et al., 2015). Here, we describe a simple and informative method to study and identify the RNA region of interaction between an RBP and its target transcript (Panda et al., 2014 and 2016). Its reproducibility and ease of use make this protocol a fast and useful method to identify interactions between RBPs and specific RNAs.

In vitro Assays for the Detection of Calreticulin Exposure, ATP and HMGB1 Release upon Cell Death

Featured protocol,  Authors: Yuting Ma
Yuting MaAffiliation 1: , Suzhou Institute of Systems Medicine, Suzhou, China
Affiliation 2: , Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China
For correspondence: yuting_ma1984@163.com
Bio-protocol author page: a3918
 and Heng Yang
Heng YangAffiliation 1: , Suzhou Institute of Systems Medicine, Suzhou, China
Affiliation 2: , Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China
Bio-protocol author page: a3919
date: 12/20/2016, 218 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2076.

Brief version appeared in Cancer Res, Sep 2015
Accumulating evidence is revealing the essential role of immune system in cancer treatment. Certain chemotherapeutic drugs can potently induce the release of ‘cell death associated molecular patterns’ (CDAMPs), which accompanies cancer cell demise. CDAMPs can engage corresponding receptors on immune cells and stimulate immune responses to achieve long-term tumor control (Ma et al., 2013; Ma et al., 2014; Yang et al., 2015). Among reported CDAMPs, calreticulin (CALR), ATP and HMGB1 are well known for their immune-stimulatory effect. Here we describe the assays that we applied to measure cell death and these CDAMPs. Briefly, cell death can be analyzed by co-staining of 4’,6-diamidino-2-phenylindole (DAPI) with 3,3’-Dihexyloxacarbocyanine Iodide [DiOC6(3)] or Annexin V. CALR exposure on the cell membrane can be detected by flow cytometry. ATP and HMGB1 release can be quantified by luminescence assay and ELISA assay respectively.

In vivo Efficacy Studies in Cell Line and Patient-derived Xenograft Mouse Models

Authors: Elizabeth A. Tovar
Elizabeth A. TovarAffiliation: Center for Cancer and Cell Biology, Van Andel Research Institute, Grand Rapids, Michigan, USA
Bio-protocol author page: a3966
Curt J. Essenburg
Curt J. EssenburgAffiliation: Center for Cancer and Cell Biology, Van Andel Research Institute, Grand Rapids, Michigan, USA
Bio-protocol author page: a3967
 and Carrie Graveel
Carrie GraveelAffiliation: Center for Cancer and Cell Biology, Van Andel Research Institute, Grand Rapids, Michigan, USA
For correspondence: carrie.graveel@vai.org
Bio-protocol author page: a3968
date: 1/5/2017, 87 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2100.

[Abstract] In vivo xenograft models derived from human cancer cells have been a gold standard for evaluating the genetic drivers of cancer and are valuable preclinical models for evaluating the efficacy of cancer therapeutics. Recently, patient-derived tumorgrafts from multiple tumor types have been developed and shown to more accurately recapitulate the molecular ...

Measuring Procaspase-8 and -10 Processing upon Apoptosis Induction

Authors: Sabine Pietkiewicz
Sabine PietkiewiczAffiliation: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Present address: Medical advisory service of social health insurance, Essen, Germany
Bio-protocol author page: a3934
Clara Wolfe
Clara WolfeAffiliation: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Bio-protocol author page: a3935
Jörn H. Buchbinder
Jörn H. BuchbinderAffiliation: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Bio-protocol author page: a3936
 and Inna N. Lavrik
Inna N. LavrikAffiliation 1: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Affiliation 2: Federal research center Institute of Cytology and Genetics, Novosibirsk, Russia
For correspondence: inna.lavrik@med.ovgu.de
Bio-protocol author page: a3937
date: 1/5/2017, 97 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2081.

[Abstract] Apoptosis or programmed cell death is important for multicellular organisms to keep cell homeostasis and for the clearance of mutated or infected cells. Apoptosis can be induced by intrinsic or extrinsic stimuli. The first event in extrinsic apoptosis is the formation of the Death-Inducing Signalling Complex (DISC), where the initiator caspases-8 and ...

Generation of Tumour-stroma Minispheroids for Drug Efficacy Testing

Authors: Mark Watters
Mark Watters Affiliation: Apoptosis Research Centre, School of Natural Sciences, National University of Ireland, Galway, Ireland
Bio-protocol author page: a3964
 and Eva Szegezdi
Eva SzegezdiAffiliation: Apoptosis Research Centre, School of Natural Sciences, National University of Ireland, Galway, Ireland
For correspondence: eva.szegezdi@nuigalway.ie
Bio-protocol author page: a3965
date: 1/5/2017, 88 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2091.

[Abstract] The three-dimensional organisation of cells in a tissue and their interaction with adjacent cells and extracellular matrix is a key determinant of cellular responses, including how tumour cells respond to stress conditions or therapeutic drugs (Elliott and Yuan, 2011). In vivo, tumour cells are embedded in a stroma formed primarily by fibroblasts that ...

Relative Stiffness Measurements of Cell-embedded Hydrogels by Shear Rheology in vitro

Authors: Thomas R. Cox
Thomas R. CoxAffiliation: The Garvan Institute of Medical Research and The Kinghorn Cancer Centre, Cancer Division, St Vincent's Clinical School, Faculty of Medicine, University of New South Wales, Sydney, Australia
For correspondence: t.cox@garvan.org.au
Bio-protocol author page: a3986
 and Chris D. Madsen
Chris D. MadsenAffiliation: Department of Laboratory Medicine, Division of Translational Cancer Research, Lund University, Lund, Sweden
For correspondence: chris.madsen@med.lu.se
Bio-protocol author page: a3987
date: 1/5/2017, 110 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2101.

[Abstract] Hydrogel systems composed of purified extracellular matrix (ECM) components (such as collagen, fibrin, Matrigel, and methylcellulose) are a mainstay of cell and molecular biology research. They are used extensively in many applications including tissue regeneration platforms, studying organ development, and pathological disease models such as cancer. ...

In vitro Histone H3 Cleavage Assay for Yeast and Chicken Liver H3 Protease

Authors: Sakshi Chauhan
Sakshi ChauhanAffiliation: Laboratory of Chromatin Biology, Department of Biological Sciences, Indian Institute of Science Education and Research, Bhopal, India
Bio-protocol author page: a3946
Gajendra Kumar Azad
Gajendra Kumar AzadAffiliation 1: Laboratory of Chromatin Biology, Department of Biological Sciences, Indian Institute of Science Education and Research, Bhopal, India
Affiliation 2: Department of Genetics, Institute of Life Sciences, Hebrew University of Jerusalem, Jerusalem, Israel
Bio-protocol author page: a3947
 and Raghuvir Singh Tomar
Raghuvir Singh TomarAffiliation: Laboratory of Chromatin Biology, Department of Biological Sciences, Indian Institute of Science Education and Research, Bhopal, India
For correspondence: rst@iiserb.ac.in
Bio-protocol author page: a3948
date: 1/5/2017, 112 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2085.

[Abstract] Histone proteins are subjected to a wide array of reversible and irreversible post-translational modifications (PTMs) (Bannister and Kouzarides, 2011; Azad and Tomar, 2014). The PTMs on histones are known to regulate chromatin structure and function. Histones are irreversibly modified by proteolytic clipping of their tail domains. The proteolytic clipping ...

Affinity Pulldown of Biotinylated RNA for Detection of Protein-RNA Complexes

Authors: Amaresh C Panda
Amaresh C PandaAffiliation: Laboratory of Genetics and Genomics, National Institute on Aging, National Institutes of Health, 251 Bayview Blvd., Baltimore, United States
For correspondence: amaresh.panda@nih.gov
Bio-protocol author page: a3875
Jennifer L. Martindale
Jennifer L. Martindale Affiliation: Laboratory of Genetics and Genomics, National Institute on Aging, National Institutes of Health, 251 Bayview Blvd., Baltimore, United States
Bio-protocol author page: a3880
 and Myriam Gorospe
Myriam GorospeAffiliation: Laboratory of Genetics and Genomics, National Institute on Aging, National Institutes of Health, 251 Bayview Blvd., Baltimore, United States
Bio-protocol author page: a3881
date: 12/20/2016, 228 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2062.

[Abstract] RNA-binding proteins (RBPs) have recently emerged as crucial players in the regulation of gene expression. The interactions of RBPs with target mRNAs control the levels of gene products by altering different regulatory steps, including pre-mRNA splicing and maturation, nuclear mRNA export, and mRNA stability and translation (Glisovic et al., 2008). ...

In vitro Assays for the Detection of Calreticulin Exposure, ATP and HMGB1 Release upon Cell Death

Authors: Yuting Ma
Yuting MaAffiliation 1: , Suzhou Institute of Systems Medicine, Suzhou, China
Affiliation 2: , Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China
For correspondence: yuting_ma1984@163.com
Bio-protocol author page: a3918
 and Heng Yang
Heng YangAffiliation 1: , Suzhou Institute of Systems Medicine, Suzhou, China
Affiliation 2: , Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China
Bio-protocol author page: a3919
date: 12/20/2016, 218 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2076.

[Abstract] Accumulating evidence is revealing the essential role of immune system in cancer treatment. Certain chemotherapeutic drugs can potently induce the release of ‘cell death associated molecular patterns’ (CDAMPs), which accompanies cancer cell demise. CDAMPs can engage corresponding receptors on immune cells and stimulate immune responses to achieve long-term ...

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, 233 views, 0 Q&A
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, 278 views, 0 Q&A
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, 352 views, 0 Q&A
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 ...
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 

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, 51658 views, 6 Q&A
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, 41555 views, 7 Q&A
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, 37164 views, 2 Q&A
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, 34791 views, 5 Q&A
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, 30713 views, 1 Q&A
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, 29955 views, 6 Q&A
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, 26034 views, 1 Q&A
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, 22867 views, 1 Q&A
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, 22520 views, 3 Q&A
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, 22262 views, 2 Q&A
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. ...
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15