University of California

Nanocav, LLC

Nantbio, Inc.

Ting-Hsiang Sherry Wu

Pei-Yu E Chiou

Michael A Teitell

Yu-Chun Kung

Yi-Chien Wu

Sung-Yong Park

Yue Chen

Kayvan R Niazi

Peter Sieling

Artin Mehrabi

Wade Nichols Richardson

Jon Thomas Van Lew

Sheraz Kalim Butt

Methods, devices, and systems are provided for the delivery of agents (e.g., nucleic acids, proteins, organic molecules, organelles, antibodies or other ligands, etc.) into live cells and/or the extraction of the same from said cells. In various embodiments the photothermal platforms and systems incorporating such photothermal platforms are provided that permit efficient, high-throughput cargo delivery into live cells.

High-throughput cargo delivery into live cells using photothermal platforms

In various embodiments, method and devices for delivering large cargos (e.g., organelles, chromosomes, bacteria, and the like) into cells are provided. In certain embodiments method of delivering a large cargo into eukaryotic cells, are provided that involve providing eukaryotic cells disposed on one side of a porous membrane; providing the cargo to be delivered in a solution disposed in a reservoir chamber on the opposite side of the porous membrane; and applying pressure to the reservoir chamber sufficient to pass the cargo through pores comprising said porous membrane wherein said cargo passes through cell membranes and into the cells.

Efficient delivery of large cargos into cells on a porous substrate

Devices and methods are presented for delivery of various macrostructures into cells, such as depleted cells as well as methods of generating engineered cells using said devices and methods. Cells are placed on a porous membrane. A force is applied by a configurable actuator to a deformable fluid reservoir that generates an applied pressure to macrostructures in a solution, causing the macrostructures to pass through the porous membrane and triggering uptake of at least some of the macrostructures into the cells to form transfected cells. Said devices and methods may be used in a process to replace defective endogenous mtDNA with corrected mtDNA to generate cellular-based therapeutics for administration to a patient. The customizable actuator may be configured with parameters to optimize efficiency for a given cell type and material to be transfected. Machine learning techniques also may be utilized to optimize transfection parameters.

Devices and methods for mitochondria replacement and for generating cellular therapeutics

Methods are provided for transfecting cells with large cargo using a poly(beta-amino ester) (PBAE) molecule, and achieving high efficiency and viability. A method is provided of transfecting cells with a cargo, by forming a complex of the cargo with a (PBAE) molecule, mixing the complex with a first buffer and contacting the complex with the cells, wherein the cargo has a dimension of at least 0.1 &#x3bc;m. The PBAE molecule may be formed by reacting an amine with a di(acrylate ester). In some aspects, the PBAE molecule is poly(1,4-butanediol diacrylate-co-4-amino-1-butanol). In some aspects, the PBAE molecule is capped with 1-(3-aminopropyl)-4-methylpiperazine.

Methods of transfection for large cargo using poly(beta-amino esters)

Methods, devices, and systems are provided for the delivery of agents (e.g., nucleic acids, proteins, organic molecules, organelles, antibodies or other ligands, 5 etc.) into live cells and/or the extraction of the same from said cells. In various embodiments the photothermal platforms and systems incorporating such photothermal platforms are provided that permit efficient, high-throughput cargo delivery into live cells.

Methods and devices for the formation and/or merging of droplets in microfluidic systems are provided. In certain embodiments a microfluidic droplet merger component is provided that comprises a central channel comprising a plurality of elements disposed and spaced to create a plurality of lateral passages that drain a carrier fluid out of a fluid stream comprising droplets of a first fluid contained in the carrier fluid; and a deformable lateral membrane valve disposed to control the width of said center channel.

High-speed on demand microfluidic droplet generation and manipulation

This invention provides novel tools for surgery on single cells and substrates/devices for delivery of reagents to selected cells. In certain embodiments the substrates comprise a surface comprising one or more orifices, where nanoparticles and/or a thin film is deposited on a surface of said orifice or near said orifice, where the nanoparticles and/or a thin film are formed of materials that heat up when contacted with electromagnetic radiation. In certain embodiments the pores are in fluid communication with microchannels containing one or more reagents to be delivered into the cells.

Photothermal substrates for selective transfection of cells

Methods and devices are provided for focusing and/or sorting activated T cells. The device comprises a microfluidic channel comprising a plurality of electrodes arranged to provide dielectrophoretic (DEP) forces that are perpendicular to forces from hydrodynamic flows along the channel. The device may be configured to apply voltages to a plurality of electrodes in a first upper region of the microfluidic channel to focus the cells into a single flow, and to apply different voltages to a plurality of electrodes in a second downstream region of the microfluidic channel to sort cells based on size. The output of the microfluidic channel may diverge into multiple channels, wherein cells of various sorted sizes are directed into the appropriate output channel.

Methods of sorting activated T cells using three dimensional dielectrophoresis

Methods and devices for the formation of droplets of a first fluid in a second fluid and the encapsulation of particles or cells within such droplets are disclosed. Impetus for droplet formation is provided by the creation of a transient bubble, which may be induced using a pulsed laser. Droplet volume and the frequency at which droplets are formed can be controlled by modulation of the pulsed laser. The disclosed methods and devices are particularly suitable for use in microfluidic devices.

High-speed on demand droplet generation and single cell encapsulation driven by induced cavitation

In certain embodiments this invention provides a pulsed-laser triggered microfluidic switching mechanism that can achieve a switching time of 70 &#x3bc;s. This switching speed is two orders of magnitude shorter than that of the fastest switching mechanism utilized in previous &#x3bc;FACS.

Growing community of inventors

Culver City, CA, United States of America

Ting-Hsiang Sherry Wu