Developing a Three-Dimensional Microenvironment to Investigate Metastatic Cancer Invasion

...................................................................................................................4 Chapter 1: Introduction..........................................................................................5 1.1 Mechanics in Cancer Metastasis....................................................................................5 1.2 Previous Works..............................................................................................................7 1.3 Motivation for Thesis.....................................................................................................9 Chapter 2: Materials and Methods......................................................................12 2.1 Fabrication of Three-Dimensional Hydrogel Scaffold................................................12 2.2 MDA-MB-231 Cell Culture.......................................................................................17 2.2.1 Labeling Cells with Fluorescent Dye............................................................18 2.3 Fabrication of Two-Dimensional Hydrogel Scaffold...................................................18 2.3.1 Chemical Activation of Coverslips...............................................................20 2.4 Cell Imaging.................................................................................................................20 2.5 Three-dimensional Traction Force Microscopy...........................................................21 Chapter 3: Results and Discussion.......................................................................22 3.1 Scaffold Results...........................................................................................................22 3.2 Calculation of Cell Tractions.......................................................................................25 3.3 Cell Traction Data on 2D Control Samples.................................................................29 Chapter 4: Future Work.......................................................................................33 4.1 Immunohistochemistry.................................................................................................33 4.2 Scaffold Improvement.................................................................................................33 4.3 Cell Tractions for 3D Hydrogel Scaffolds...................................................................35 References...............................................................................................................36