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Cancer Biology

Glioma Induction by Intracerebral Retrovirus Injection

Featured protocol,  Authors: Ravinder K Verma
Ravinder K Verma Affiliation: Division of Experimental Hematology and Cancer Biology, Brain Tumor Center, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, USA
Bio-protocol author page: a4878
Fanghui Lu
Fanghui LuAffiliation: Division of Experimental Hematology and Cancer Biology, Brain Tumor Center, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, USA
Bio-protocol author page: a4879
 and Qing Richard Lu
Qing Richard LuAffiliation: Division of Experimental Hematology and Cancer Biology, Brain Tumor Center, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, USA
For correspondence: richard.lu@cchmc.org
Bio-protocol author page: a4880
date: 7/20/2017, 22 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2404.

Brief version appeared in Cancer Cell, May 2016
Glioblastoma (GBM) is the most common primary brain cancer in adults and has a poor prognosis. It is characterized by a high degree of cellular infiltration that leads to tumor recurrence, atypical hyperplasia, necrosis, and angiogenesis. Despite aggressive treatment modalities, current therapies are ineffective for GBM. Mouse GBM models not only provide a better understanding in the mechanisms of gliomagenesis, but also facilitate the drug discovery for treating this deadly cancer. A retroviral vector system that expresses PDGFBB (Platelet-derived growth factor BB) and inactivates PTEN (Phosphatase and tensin homolog) and P53 tumor suppressors provides a rapid and efficient induction of glioma in mice with full penetrance. In this protocol, we describe a simple and practical method for inducing GBM formation by retrovirus injection in the murine brain. This system gives a spatial and temporal control over the induction of glioma and allows the assessment of therapeutic effects with a bioluminescent reporter.

Non-radioactive LATS in vitro Kinase Assay

Featured protocol,  Authors: Audrey W. Hong
Audrey W. HongAffiliation: Department of Pharmacology and Moores Cancer Center, University of California San Diego, La Jolla, California 92093, USA
Bio-protocol author page: a4827
 and Kun-Liang Guan
Kun-Liang GuanAffiliation: Department of Pharmacology and Moores Cancer Center, University of California San Diego, La Jolla, California 92093, USA
For correspondence: kuguan@ucsd.edu
Bio-protocol author page: a868
date: 7/20/2017, 15 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2391.

Brief version appeared in EMBO Rep, Jan 2017
This protocol describes a method to directly measure LATS activity by an in vitro kinase assay using YAP as a substrate.

Analyzing the Properties of Murine Intestinal Mucins by Electrophoresis and Histology

Featured protocol,  Authors: Ran Wang
Ran WangAffiliation: Inflammatory Disease Biology and Therapeutics Group, Mater Research Institute–The University of Queensland, Translational Research Institute, Brisbane, Australia
Bio-protocol author page: a4833
 and Sumaira Z. Hasnain
Sumaira Z. HasnainAffiliation: Inflammatory Disease Biology and Therapeutics Group, Mater Research Institute–The University of Queensland, Translational Research Institute, Brisbane, Australia
For correspondence: sumaira.hasnain@mater.uq.edu.au
Bio-protocol author page: a4834
date: 7/20/2017, 20 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2394.

Brief version appeared in PLoS Pathog, Feb 2017
Specialized secretory cells known as goblet cells in the intestine and respiratory epithelium are responsible for the secretion of mucins. Mucins are large heavily glycosylated proteins and typically have a molecular mass higher than 106 Da. These large proteins are densely substituted with short glycan chains, which have many important functional roles including determining the hydration and viscoelastic properties of the mucus gel that lines and protects the intestinal epithelium. In this protocol, we comprehensively describe the method for extraction of murine mucus and its analysis by agarose gel electrophoresis. Additionally we describe the use of High Iron Diamine-Alcian Blue, Periodic Acid Schiff’s-Alcian Blue and immune–staining methods to identify and differentiate between the different states of glycosylation on these mucin glycoproteins, in particular with a focus on sulphation and sialylation.

Measurement of the Intracellular Calcium Concentration with Fura-2 AM Using a Fluorescence Plate Reader

Featured protocol,  Authors: Magdiel Martínez
Magdiel MartínezAffiliation: Department of Physiology and Biophysics, Medical Sciences Campus, University of Puerto Rico, San Juan, Puerto Rico
Bio-protocol author page: a4908
Namyr A. Martínez
Namyr A. MartínezAffiliation: Department of Physiology and Biophysics, Medical Sciences Campus, University of Puerto Rico, San Juan, Puerto Rico
Bio-protocol author page: a4909
 and Walter I. Silva
Walter I. SilvaAffiliation: Department of Physiology and Biophysics, Medical Sciences Campus, University of Puerto Rico, San Juan, Puerto Rico
For correspondence: walter.silva@upr.edu
Bio-protocol author page: a4910
date: 7/20/2017, 19 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2411.

Brief version appeared in J Biol Chem, Jun 2016
Intracellular calcium elevation triggers a wide range of cellular responses. Calcium responses can be affected or modulated by membrane receptors mutations, localization, exposure to agonists/antagonists, among others (Burgos et al., 2007; Martínez et al., 2016). Changes in intracellular calcium concentration can be measured using the calcium sensitive fluorescent ratiometric dye fura-2 AM. This method is a high throughput way to measure agonist mediated calcium responses.

Generation of a Cellular Reporter for Functional BRD4 Inhibition

Featured protocol,  Authors: Sara Sdelci
Sara SdelciAffiliation: CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Lazarettgasse 14, Vienna, Austria
Bio-protocol author page: a4753
 and Stefan Kubicek
Stefan KubicekAffiliation 1: CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Lazarettgasse 14, Vienna, Austria
Affiliation 2: Christian Doppler Laboratory for Chemical Epigenetics and Antiinfectives, CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria
For correspondence: skubicek@cemm.oeaw.ac.at
Bio-protocol author page: a4754
date: 7/5/2017, 177 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2368.

Brief version appeared in Nat Chem Biol, Jul 2016
The ubiquitously expressed bromodomain-containing protein 4 (BRD4) is an epigenetic reader, which recruits transcriptional regulatory complexes to acetylated chromatin. Because of its role in enhancing proliferation, BRD4 has become a therapeutic target in oncology, as the inhibition of this protein leads to the reduction of the growth of many tumours. Even though BRD4 is more and more studied, its mechanism of action has not been fully described yet. Therefore, we aimed at generating a cellular reporter system to monitor BRD4 inhibition. Such reporter can be potentially used in high throughput chemical and genetic screenings, in order to uncover new possible BRD4 functional pathways. The deeper understanding of the mechanism of action of BRD4 activity will certainly help in developing new therapy strategies for those cancers so called BRD4-dependent.

Tumorigenicity Assay in Nude Mice

Featured protocol,  Authors: Feng Du
Feng DuAffiliation: State key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi'an, China
Bio-protocol author page: a4714
Xiaodi Zhao
Xiaodi ZhaoAffiliation: State key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi'an, China
For correspondence: leedyzhao@fmmu.edu.cn
Bio-protocol author page: a4715
 and Daiming Fan
Daiming FanAffiliation: State key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, China
Bio-protocol author page: a4716
date: 7/5/2017, 178 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2364.

Brief version appeared in J Cell Biol, Aug 2015
Tumorigenicity refers to the ability of cultured cells to develop viable tumors in immune-deficient animals. The goal of this protocol is to illustrate tumorigenicity assay by subcutaneous tumor-cell-transplantation in nude mice. Target cells are transplanted to 6-week-old nude mice subcutaneously and the tumor growth is monitored over a period of observation or treatment. When tumor grows to a pre-determined size or by the end of the limited period, the nude mice will be euthanatized and the xenograft will removed for further examination.

A Novel Mouse Skin Graft Model of Vascular Tumors Driven by Akt1

Featured protocol,  Authors: Thuy L. Phung
Thuy L. PhungAffiliation: Department of Pathology, Texas Children’s Hospital and Baylor College of Medicine, Houston, USA
For correspondence: tphung@bcm.edu
Bio-protocol author page: a4800
 and Sriram Ayyaswamy
Sriram AyyaswamyAffiliation: Department of Pathology, Texas Children’s Hospital and Baylor College of Medicine, Houston, USA
Bio-protocol author page: a4801
date: 7/5/2017, 183 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2369.

Brief version appeared in Cancer Res, Jan 2015
To investigate whether endothelial Akt1 activation is sufficient to induce vascular tumor formation in the skin, we have developed a skin graft model in which a skin fragment from transgenic donor mice with inducible and endothelial cell-specific overexpression of activated Akt1 (myrAkt1) is grafted into the skin of wild type recipient mice. The donor skin successfully engrafts after two weeks and, more importantly, vascular tumor develops at the site of transgenic skin graft when myrAkt1 expression is turned on. This skin graft model is a novel approach to investigate the biological impact of a key signal transduction molecule in a temporal, localized and organ-specific manner.

Glioma Induction by Intracerebral Retrovirus Injection

Authors: Ravinder K Verma
Ravinder K Verma Affiliation: Division of Experimental Hematology and Cancer Biology, Brain Tumor Center, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, USA
Bio-protocol author page: a4878
Fanghui Lu
Fanghui LuAffiliation: Division of Experimental Hematology and Cancer Biology, Brain Tumor Center, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, USA
Bio-protocol author page: a4879
 and Qing Richard Lu
Qing Richard LuAffiliation: Division of Experimental Hematology and Cancer Biology, Brain Tumor Center, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, USA
For correspondence: richard.lu@cchmc.org
Bio-protocol author page: a4880
date: 7/20/2017, 22 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2404.

[Abstract] Glioblastoma (GBM) is the most common primary brain cancer in adults and has a poor prognosis. It is characterized by a high degree of cellular infiltration that leads to tumor recurrence, atypical hyperplasia, necrosis, and angiogenesis. Despite aggressive treatment modalities, current therapies are ineffective for GBM. Mouse GBM models not only provide ...

Non-radioactive LATS in vitro Kinase Assay

Authors: Audrey W. Hong
Audrey W. HongAffiliation: Department of Pharmacology and Moores Cancer Center, University of California San Diego, La Jolla, California 92093, USA
Bio-protocol author page: a4827
 and Kun-Liang Guan
Kun-Liang GuanAffiliation: Department of Pharmacology and Moores Cancer Center, University of California San Diego, La Jolla, California 92093, USA
For correspondence: kuguan@ucsd.edu
Bio-protocol author page: a868
date: 7/20/2017, 15 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2391.

[Abstract] This protocol describes a method to directly measure LATS activity by an in vitro kinase assay using YAP as a substrate....

Analyzing the Properties of Murine Intestinal Mucins by Electrophoresis and Histology

Authors: Ran Wang
Ran WangAffiliation: Inflammatory Disease Biology and Therapeutics Group, Mater Research Institute–The University of Queensland, Translational Research Institute, Brisbane, Australia
Bio-protocol author page: a4833
 and Sumaira Z. Hasnain
Sumaira Z. HasnainAffiliation: Inflammatory Disease Biology and Therapeutics Group, Mater Research Institute–The University of Queensland, Translational Research Institute, Brisbane, Australia
For correspondence: sumaira.hasnain@mater.uq.edu.au
Bio-protocol author page: a4834
date: 7/20/2017, 20 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2394.

[Abstract] Specialized secretory cells known as goblet cells in the intestine and respiratory epithelium are responsible for the secretion of mucins. Mucins are large heavily glycosylated proteins and typically have a molecular mass higher than 106 Da. These large proteins are densely substituted with short glycan chains, which have many important functional ...

Measurement of the Intracellular Calcium Concentration with Fura-2 AM Using a Fluorescence Plate Reader

Authors: Magdiel Martínez
Magdiel MartínezAffiliation: Department of Physiology and Biophysics, Medical Sciences Campus, University of Puerto Rico, San Juan, Puerto Rico
Bio-protocol author page: a4908
Namyr A. Martínez
Namyr A. MartínezAffiliation: Department of Physiology and Biophysics, Medical Sciences Campus, University of Puerto Rico, San Juan, Puerto Rico
Bio-protocol author page: a4909
 and Walter I. Silva
Walter I. SilvaAffiliation: Department of Physiology and Biophysics, Medical Sciences Campus, University of Puerto Rico, San Juan, Puerto Rico
For correspondence: walter.silva@upr.edu
Bio-protocol author page: a4910
date: 7/20/2017, 19 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2411.

[Abstract] Intracellular calcium elevation triggers a wide range of cellular responses. Calcium responses can be affected or modulated by membrane receptors mutations, localization, exposure to agonists/antagonists, among others (Burgos et al., 2007; Martínez et al., 2016). Changes in intracellular calcium concentration can be measured using the calcium sensitive ...

Generation of a Cellular Reporter for Functional BRD4 Inhibition

Authors: Sara Sdelci
Sara SdelciAffiliation: CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Lazarettgasse 14, Vienna, Austria
Bio-protocol author page: a4753
 and Stefan Kubicek
Stefan KubicekAffiliation 1: CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Lazarettgasse 14, Vienna, Austria
Affiliation 2: Christian Doppler Laboratory for Chemical Epigenetics and Antiinfectives, CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria
For correspondence: skubicek@cemm.oeaw.ac.at
Bio-protocol author page: a4754
date: 7/5/2017, 177 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2368.

[Abstract] The ubiquitously expressed bromodomain-containing protein 4 (BRD4) is an epigenetic reader, which recruits transcriptional regulatory complexes to acetylated chromatin. Because of its role in enhancing proliferation, BRD4 has become a therapeutic target in oncology, as the inhibition of this protein leads to the reduction of the growth of many tumours. ...

Tumorigenicity Assay in Nude Mice

Authors: Feng Du
Feng DuAffiliation: State key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi'an, China
Bio-protocol author page: a4714
Xiaodi Zhao
Xiaodi ZhaoAffiliation: State key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi'an, China
For correspondence: leedyzhao@fmmu.edu.cn
Bio-protocol author page: a4715
 and Daiming Fan
Daiming FanAffiliation: State key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, China
Bio-protocol author page: a4716
date: 7/5/2017, 178 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2364.

[Abstract] Tumorigenicity refers to the ability of cultured cells to develop viable tumors in immune-deficient animals. The goal of this protocol is to illustrate tumorigenicity assay by subcutaneous tumor-cell-transplantation in nude mice. Target cells are transplanted to 6-week-old nude mice subcutaneously and the tumor growth is monitored over a period of ...

A Novel Mouse Skin Graft Model of Vascular Tumors Driven by Akt1

Authors: Thuy L. Phung
Thuy L. PhungAffiliation: Department of Pathology, Texas Children’s Hospital and Baylor College of Medicine, Houston, USA
For correspondence: tphung@bcm.edu
Bio-protocol author page: a4800
 and Sriram Ayyaswamy
Sriram AyyaswamyAffiliation: Department of Pathology, Texas Children’s Hospital and Baylor College of Medicine, Houston, USA
Bio-protocol author page: a4801
date: 7/5/2017, 183 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2369.

[Abstract] To investigate whether endothelial Akt1 activation is sufficient to induce vascular tumor formation in the skin, we have developed a skin graft model in which a skin fragment from transgenic donor mice with inducible and endothelial cell-specific overexpression of activated Akt1 (myrAkt1) is grafted into the skin of wild type recipient mice. The donor ...

Soft Agar Colony Formation Assay as a Hallmark of Carcinogenesis

Authors: Feng Du
Feng DuAffiliation: State key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, China
Bio-protocol author page: a4714
Xiaodi Zhao
Xiaodi ZhaoAffiliation: State key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, China
For correspondence: leedyzhao@fmmu.edu.cn
Bio-protocol author page: a4715
 and Daiming Fan
Daiming FanAffiliation: State key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, China
Bio-protocol author page: a4716
date: 6/20/2017, 320 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2351.

[Abstract] Soft agar colony formation assay is established to estimate the anchorage-independent growth ability of cells. In this assay, a bottom layer of agar with complete media is poured and solidified first, followed by an upper layer containing a specified number of cells suspended in medium-agar mixture. After two weeks of incubation, the number of colonies ...

Targeted Nucleotide Substitution in Mammalian Cell by Target-AID

Authors: Takayuki Arazoe
Takayuki ArazoeAffiliation: Graduate school of Science, Technology and Innovation, Kobe University, Hyogo, Japan
Bio-protocol author page: a4638
Keiji Nishida
Keiji NishidaAffiliation: Graduate school of Science, Technology and Innovation, Kobe University, Hyogo, Japan
For correspondence: keiji_nishida@people.kobe-u.ac.jp
Bio-protocol author page: a4639
 and Akihiko Kondo
Akihiko KondoAffiliation: Graduate school of Science, Technology and Innovation, Kobe University, Hyogo, Japan
For correspondence: akondo@kobe-u.ac.jp
Bio-protocol author page: a1574
date: 6/5/2017, 453 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2339.

[Abstract] Programmable RNA-guided nucleases based on CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated protein) systems has been applied to various type of cells as powerful genome editing tools. By using activation-induced cytidine deaminase (AID) in place of the nuclease activity of the CRISPR/Cas9 system, we have developed ...

Intracaecal Orthotopic Colorectal Cancer Xenograft Mouse Model

Authors: Hsin-Wei Liao
Hsin-Wei LiaoAffiliation 1: Department of Molecular and Cellular Oncology, the University of Texas MD Anderson Cancer Center, Houston, USA
Affiliation 2: Center for Systems Biology, Massachusetts General Hospital Research Institute and Harvard Medical School, Boston, USA
Bio-protocol author page: a4595
 and Mien-Chie Hung
Mien-Chie HungAffiliation 1: Department of Molecular and Cellular Oncology, the University of Texas MD Anderson Cancer Center, Houston, USA
Affiliation 2: Graduate Institute of Biomedical Sciences and Center for Molecular Medicine, a Medical University, Taichung, Taiwan
Affiliation 3: Department of Biotechnology, Asia University, Taichung, Taiwan
For correspondence: mhung@mdanderson.org
Bio-protocol author page: a886
date: 6/5/2017, 337 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2311.

[Abstract] The host microenvironment plays a prominent role in tumor growth, angiogenesis, invasion, metastasis, and response to therapy. Orthotopic tumor model mimics the natural environment of tumor development and provides an effective approach to investigate tumor pathophysiology and develop therapeutic strategies. This protocol describes the technique involving ...
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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, 57761 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, 44518 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, ...

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, 40781 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 ...

[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, 40707 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 ...

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, 34088 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, 32517 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, 30364 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, 26785 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, 26000 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, 24753 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. ...
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