February 2012: This Month in JoVE

JoVE 4258 2/01/2012

Aaron Kolski-Andreaco

Microfluidic Device for Recreating a Tumor Microenvironment in Vitro

JoVE 2425 11/20/2011

Bhushan J. Toley*, Dan E. Ganz*, Colin L. Walsh, Neil S. Forbes

Department of Chemical Engineering, University Of Massachusetts Amherst

We present the procedure for fabrication and operation of a microfluidic device that recreates heterogeneous tumor microenvironments in vitro. The variability in apoptosis within tumor tissue was quantified using fluorescent stains and the effective diffusion coefficient of the chemotherapeutic drug doxorubicin into tumor tissue was evaluated.

An Orthotopic Model of Serous Ovarian Cancer in Immunocompetent Mice for in vivo Tumor Imaging and Monitoring of Tumor Immune Responses

JoVE 2146 11/28/2010

Selene Nunez-Cruz¹, Denise C. Connolly², Nathalie Scholler¹

¹Penn Ovarian Cancer Research Center, Center for Research on Reproduction and Womans Health, Department of Obstetrics and Gynecology, University of Pennsylvania-School of Medicine, ²Women's Cancer Program, Fox Chase Cancer Center

To study in vivo tumor growth and tumor microenvironment, we used a syngeneic and orthotopic mouse model of ovarian cancer in immunocompetent animals. We transduced a mouse tumor cell line (MOV1) with Katushka fluorescent protein (MOV1^KAT) and here we show its orthotopic implantation in ovary and in vivo imaging.

MAME Models for 4D Live-cell Imaging of Tumor: Microenvironment Interactions that Impact Malignant Progression

JoVE 3661 2/17/2012

Mansoureh Sameni¹, Arulselvi Anbalagan¹, Mary B. Olive¹, Kamiar Moin^1,2, Raymond R. Mattingly^1,2, Bonnie F. Sloane^1,2

¹Department of Pharmacology, Wayne State University , ²Barbara Ann Karmanos Cancer Institute, Wayne State University

We have developed 3D coculture models for live-cell imaging in real-time of interactions among breast tumor cells and other cells in their microenvironment that impact progression to an invasive phenotype. These models can serve as preclinical screens for drugs to target paracrine-induced proteolytic, chemokine/cytokine and kinase pathways implicated in invasiveness.

Dendra2 Photoswitching through the Mammary Imaging Window

JoVE 1278 6/05/2009

Bojana Gligorijevic ^1,2, Dmitriy Kedrin¹, Jeffrey E Segall¹, John Condeelis^1,2, Jacco van Rheenen^1,2,3

¹Department of Anatomy and Structural Biology, Albert Einstein College of Medicine - Yeshiva University, ²Gruss Lipper Biophotonics Center, Albert Einstein College of Medicine - Yeshiva University, ³Hubrecht Institute-KNAW and University Medical Center Utrecht

Intravital photoswitching and tracking of Dendra2-labeled tumor cells through the Mammary Imaging Window is a technique which allows us to image the metastatic behavior of tumor cells in chosen tumor microenvironments over a timescale of days.

Isolation of Mammary Epithelial Cells from Three-dimensional Mixed-cell Spheroid Co-culture

JoVE 3760 4/30/2012

Kun Xu, Rachel J. Buchsbaum

Molecular Oncology Research Institute, Department of Medicine, Tufts Medical Center

A simple method is described for analyzing effects of tissue fibroblasts on associated epithelial cells. The combination of this method and three-dimensional tissue culture can facilitate analysis of cells after isolation from 3D. The technique is applicable to cells of varying malignant potential, allowing systematic study of effects of tumor-associated stroma on tumor cells.

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

JoVE 2941 7/25/2011

Amanda Gatesman Ammer¹, Karen E. Hayes¹, Karen H. Martin¹, Lingqing Zhang², George A. Spirou³, Scott A. Weed¹

¹Department of Neurobiology and Anatomy, Program in Cancer Cell Biology, Mary Babb Randolph Cancer Center, West Virginia University, ²Sensory Neuroscience Research Center, West Virginia University , ³Departments of Otolaryngology and Physiology, Center for Neuroscience, West Virginia University

A comprehensive overview of the techniques involved in generating a mouse model of oral cancer and quantitative monitoring of tumor invasion within the tongue through multi-photon microscopy of labeled cells is presented. This system can serve as a useful platform for the molecular assessment and drug efficacy of anti-invasive compounds.

Quantitative Analysis of Cancer Metastasis using an Avian Embryo Model

JoVE 2815 5/30/2011

Trenis D. Palmer¹, John Lewis², Andries Zijlstra¹

¹Department of Pathology, Vanderbilt University , ²Departments of Oncology, Surgery and Medical Biophysics, London Regional cancer program

Using quantitative PCR, we demonstrate how the well-established chick CAM model can be used to quantitatively analyze the metastasis of human tumor cells to distant organs.

Detection of Functional Matrix Metalloproteinases by Zymography

JoVE 2445 11/08/2010

Xueyou Hu, Christine Beeton

Department of Molecular Physiology and Biophysics, Baylor College of Medicine

This protocol describes an activity-based assay for detecting matrix metalloproteinases in culture supernatants or body fluids.

Micropatterned Surfaces to Study Hyaluronic Acid Interactions with Cancer Cells

JoVE 2413 12/22/2010

Laura E. Dickinson, Sharon Gerecht

Department of Chemical and Biomolecular Engineering, Johns Hopkins Physical Sciences Oncology Center and Institute for NanoBioTechnology, Johns Hopkins University

A novel approach that allows the high-resolution analysis of cancer cell interactions with exogenous hyaluronic acid (HA) is described. Patterned surfaces are fabricated by combining carbodiimide chemistry and microcontact printing.