Spheroid Assay to Measure TGF-β-induced Invasion

JoVE 3337 11/16/2011

Hildegonda P.H. Naber, Eliza Wiercinska, Peter ten Dijke, Theo van Laar

Department of Molecular Cell Biology and Centre for Biomedial Genetics, Leiden University Medical Centre

An assay to quantitatively measure Transforming Growth Factor (TGF)-β-induced invasion in 3-dimensional collagen gels is described. This assay takes advantage of the MCF10A series of cell lines, which represent different stages of breast cancer development. This method can be adopted to be used with other cell lines and might be used to investigate other potential activators or inhibitors of invasion.

In vitro Mesothelial Clearance Assay that Models the Early Steps of Ovarian Cancer Metastasis

JoVE 3888 2/17/2012

Rachel A. Davidowitz, Marcin P. Iwanicki, Joan S. Brugge

Department of Cell Biology, Harvard Medical School

The mesothelial clearance assay described here takes advantage of fluorescently labeled cells and time-lapse video microscopy to visualize and quantitatively measure the interactions of ovarian cancer multicellular spheroids and mesothelial cell monolayers. This assay models the early steps of ovarian cancer metastasis.

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.

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.

A Simple Hanging Drop Cell Culture Protocol for Generation of 3D Spheroids

JoVE 2720 5/06/2011

Ramsey Foty

Department of Surgery, UMDNJ-Robert Wood Johnson Medical School

We describe a simple, rapid method of generating 3D tissue-like spheroids and their potential application to quantify differences in cell-cell interactions.

Isolation of Retinal Stem Cells from the Mouse Eye

JoVE 2209 9/11/2010

Brenda L.K. Coles, Derek van der Kooy

Molecular Genetics, University of Toronto

In this video, we will demonstrate how to isolate retinal stem cells from the ciliary epithelium of the mouse eye and grow them in culture to form clonal retinal spheres. The spheres that are isolated possess the cardinal properties of stem cells: self-renewal and multipotentiality.

Measurement of Aggregate Cohesion by Tissue Surface Tensiometry

JoVE 2739 4/08/2011

Christine M. Butler, Ramsey A. Foty

Department of Surgery, UMDNJ-Robert Wood Johnson Medical School

We describe a method of measuring binding energy, expressible as tissue surface tension, between cells within 3D tissue-like aggregates. Differences in tissue surface tension have been demonstrated to correlate with invasiveness of lung, muscle, and brain tumors, and are fundamental determinants of establishing spatial relationships between different cell types.

Establishment and Propagation of Human Retinoblastoma Tumors in Immune Deficient Mice

JoVE 2644 8/04/2011

Wesley S. Bond^1,2, Lalita Wadhwa^2,3, Laszlo Perlaky^2,3, Rebecca L. Penland⁴, Mary Y. Hurwitz^2,3, Richard L. Hurwitz*^2,3,5,7, Patricia Chèvez-Barrios*^4,5,6

¹Interdepartmental Program in Translational Biology & Molecular Medicine, Baylor College of Medicine, ²Texas Children's Cancer Center, Baylor College of Medicine, ³Department of Pediatrics, Baylor College of Medicine, ⁴Department of Pathology, The Methodist Hospital Research Institute, ⁵Department of Ophthalmology, Retinoblastoma Center of Houston, ⁶Center for Cell and Gene Therapy, Baylor College of Medicine, ⁷Baylor College of Medicine, Center for Cell and Gene Therapy

A method is described to propagate human retinoblastoma tumors in mice. Tumor cells are directly injected into the eyes of immune deficient mice. Secondary tumors have been successfully established using both cells directly harvested from human tumors and cultured tumorspheres.

February 2012: This Month in JoVE

JoVE 4258 2/01/2012

Aaron Kolski-Andreaco

Microarray Analysis for Saccharomyces cerevisiae

JoVE 2585 4/07/2011

Scott Tighe, Tim Hunter, Pat Reed, Janet Murray

Vermont Genetics Network, The University of Vermont

In this protocol, gene expression in yeast (Saccharomyces cerevisiae) is changed after exposure to oxidative stress induced by the addition of hydrogen peroxide (H₂O₂), an oxidizing agent.