University of California, Los Angeles,
Summary:Enabling technologies for transferring large-sized cargo into mammalian cells are needed to advance key applications in cell engineering. High-throughput delivery of organelles, modified intracellular pathogens, proteins, and other cargo is required to obtain reliable statistical data. However, reliable methodologies for introducing large-sized cargo into mammalian cells at high throughput have not existed. Here, we report a massively parallel platform for high-throughput delivery of large cargo directly into the cytosol of mammalian cells. Cargo up to a micron in size or more can be reproducibly delivered into 100,000 cells on the platform in a minute. The delivery platform is a compact chip on which hundreds of thousands of micron-sized cavitation bubbles explode in response to laser pulse illumination. High-speed fluid flows near cavitation bubbles disrupt contacting cell membranes with precision, resulting in micron-sized transient membrane pores. Pressured flow provides an active driving force to speed slow diffusing large cargo through these pores before they reseal. We have reproducibly delivered large cargo including micron-sized bacteria, enzymes, antibodies, and functional nanoparticles into a variety of cell lines, including three different types of primary cells, with high efficiency and high cell viability. Massively parallel and nearly simultaneous delivery of cargo into cells under the same physiological conditions enables reliable statistical measurements of cargo interactions with cells over time.