Decoupling the Influence of Cell Proliferation and Migration in Wound Healing Assays by Combining Laser Direct Write Photoablation and Mathematical Modeling

Abstract

<p>Traditional in vitro scratch assays lack standardization due to poor control over wound geometry and fail to account for cell proliferation. Here, we developed a novel scratch assay that enables precise control over wound geometry using CAD/CAM laser photoablation and takes cell proliferation into consideration using a simple reaction-diffusion based mathematical model. We demonstrated that diffusivity in precisely photoablated cell layers serves as a more accurate measure of cell motility than the rate of gap closure. Further, we biologically validated this assay using cells harvested from patients and patient-derived xenografts to gain insights into the influence of the presence stromal cells on metaplastic and non-metaplastic triple negative breast cancer metastasis.</p>