Pilot Project 2

Revealing Mechanisms of Microvesicle Biogenesis in Breast Cancer via in situ Microscopy

Project Investigators: Lara A. Estroff  and Lena Kourkoutis

Additional Collaborators: Claudia Fischbach and Matt Paszek

This project tackles questions related to the formation mechanisms, microstructural evolution, and chemical composition of MVs produced by breast cancer cells in 2-D and 3-D culture models. To answer these questions, we are pursuing two parallel aims. Aim 1. High resolution structural and chemical characterization of MVs using cryoEM. In this aim, we are culturing MDA-MB-231 human breast cancer cells in 2D using conditions that are known to induce the production of MVs. Samples are then vitrified and prepared for high resolution imaging of the MVs by cryoTEM. Aim 2. Dynamic tracking of MV formation as a function of substrate properties. In this aim, we will use real-time confocal fluorescence microscopy to monitor, in real-time, the production of MVs as a function of substrate chemistry and mechanical properties in 2D cultures.