This invention is an intracellular microfluidic delivery device that is capable of delivering molecules across the cell membrane. This device is useful as a versatile and a widely applicable laboratory tool to deliver desired molecules into target cells.
Many pharmacologic therapeutics are small-molecule drugs, as they are able to cross selectively permeable cell membranes with relative ease. The next generation of protein, and nucleic-acid therapeutics cannot readily cross the cell membrane and thus require cellular modification to facilitate delivery. However, existing techniques for intracellular delivery to primary cells have limitations. Electroporation results in considerable cellular toxicity. Viral vectors are unable to infect resting lymphocytes. Antibody-drug complexes and conjugates require specific antibodies for each cell type and distinct designs to carry different payloads. Moreover, many clinically important cell types, such as stem cells and immune cells, are not properly addressed by existing methods. Intracellular delivery is a long-standing challenge in research and therapeutic applications, and development of systems that can deliver cargo to a variety of cell types is necessary.
Apart from fundamental research, effective intracellular delivery is important for cell engineering applications in medicine, as highlighted by its critical role in next generation therapeutics. The inventors have developed a microfluidic device that causes perturbations in the cell membrane that allow for intracellular delivery of macromolecules, such as proteins, DNA, and RNA. The microfluidic device is comprised of a flow channel containing cells in the fluid and a membrane perturbing surface. To achieve membrane-perturbation, sharp nanostructured elements can be incorporated on the surface of microfluidic devices. Collision of the cells with nanostructured microteeth, nanoteeth, spikes, and rough abrasions can create membrane perturbation for intracellular delivery. Membrane perturbing features can also include immobilized detergents, chemical compounds, special materials, and dangling chemical moieties. Effective intracellular delivery can be achieved by modulating the physical and chemical properties of the surface that cells come in contact with as they pass through the microfluidic flow channel. To help ensure contact between the perturbing features and the cells, the cells can be expanded through the use of a hypotonic buffer. The desired molecules can diffuse into the cell while perturbation of the cell membrane is occurring or after active perturbation has ended, and before the holes in the membrane close. This approach can be applied to ex vivo treatment of patient cells in therapies targeted at different disorders, such as hematological diseases.
- Intracellular delivery tool that causes increased membrane permeability without excessive cell damage
- Effectively deliver macromolecules, such as proteins, and nucleic-acid therapeutics, into the cell