Design of a microfluidic device to quantify dynamic intra-nuclear deformation during cell migration through confining environments
Year: 2015
Type: article
Abstract: The ability of cells to migrate through tissues and interstitial spaces is an essential factor during development and tissue homeostasis, immune cell mobility, and in various human diseases. Deformation of the nucleus and its associated lamina during 3-D migration is gathering increasing interest in the context of cancer metastasis, with the underlying hypothesis that a softer nucleus, resulting from reduced levels of lamin A/C, may aid tumour spreading. However, current methods to study the migration of cells in confining three dimensional (3-D) environments are limited by their imprecise control over the confinement, physiological relevance, and/or compatibility with high resolution imaging techniques. We describe the design of a polydimethylsiloxane (PDMS) microfluidic device composed of channels with precisely-defined constrictions mimicking physiological environments that enable high resolution imaging of live and fixed cells. The device promotes easy cell loading and rapid, yet long-lasting (>24 hours) chemotactic gradient formation without the need for continuous perfusion. Using this device, we obtained detailed, quantitative measurements of dynamic nuclear deformation as cells migrate through tight spaces, revealing distinct phases of nuclear translocation through the constriction, buckling of the nuclear lamina, and severe intranuclear strain. Furthermore, we found that lamin A/C-deficient cells exhibited increased and more plastic nuclear deformations compared to wild-type cells but only minimal changes in nuclear volume, implying that low lamin A/C levels facilitate migration through constrictions by increasing nuclear deformability rather than compressibility. The integration of our migration devices with high resolution time-lapse imaging provides a powerful new approach to study intracellular mechanics and dynamics in a variety of physiologically-relevant applications, ranging from cancer cell invasion to immune cell recruitment.
Source: Integrative Biology
DOI: 10.1039/c5ib00200a
Institution: Cornell University
Language: English
Subfield: Cell Biology
Domain: Life Sciences
Open Access status: bronze
Funders: National Science Foundation, U.S. Department of Defense, National Institutes of Health, Directorate for Biological Sciences, National Heart, Lung, and Blood Institute, National Cancer Institute, National Institute of Neurological Disorders and Stroke, Division of Chemical, Bioengineering, Environmental, and Transport Systems, Division of Electrical, Communications and Cyber Systems, Air Force Civil Engineer Center
Awards: CAREER:The role of nuclear biomechanics during cell migration in 3-D environments, G1957246597, G2519517127, Nuclear mechanics and mechanotransduction in muscular laminopathies, G4645538493, G4715424683, NNIN: National Nanotechnology Infrastructure Network, G638204625, G6671297155, G6851509504, G7965920530, G8550167110, G9000148656, G9000244162
OpenAlex ID: W2106226359