Active loading of extracellular vesicles (EVs) with an exogenous molecule without damaging the extracellular vesicles is provided. A composition comprising a population of extracellular vesicles loaded with an exogenous molecule, which have maintained their integrity, original endogenous cargo and functionality, compared to unloaded controls is also provided. Extracellular vesicles are derived from a stem cell, preferably an adult stem cell, or from a biological fluid, a conditioned cell medium or a tissue culture medium.

Delivering a payload across a plasma membrane of a cell includes providing a population of cells and contacting the population of cells with a volume of an aqueous solution. The aqueous solution includes the payload and alcohol content greater than 5 percent concentration. The volume of the aqueous solution may be a function of exposed surface area of the population of cells, or may be a function of a number of cells in the population of cells. Related compositions, apparatus, systems, techniques, and articles are also described.

Delivering a payload across a plasma membrane of a cell includes providing a population of cells and contacting the population of cells with a volume of an aqueous solution. The aqueous solution includes the payload and alcohol content greater than 5 percent concentration. The volume of the aqueous solution may be a function of exposed surface area of the population of cells, or may be a function of a number of cells in the population of cells. Related compositions, apparatus, systems, techniques, and articles are also described.

Delivering a payload across a plasma membrane of a cell includes providing a population of cells and contacting the population of cells with a volume of an aqueous solution. The aqueous solution includes the payload and alcohol content greater than 5 percent concentration. The volume of the aqueous solution may be a function of exposed surface area of the population of cells, or may be a function of a number of cells in the population of cells. Related compositions, apparatus, systems, techniques, and articles are also described.

Provided are a microfluidic apparatus, a method and system for introducing a substance into a cell. The microfluidic apparatus includes a cavity channel, a bulk wave generating device and a surface acoustic wave generating device; a microstructure is arranged on an inner wall of the cavity channel, and the microstructure is constructed for forming a bubble by a solution at the microstructure when the solution is injected into the cavity channel; the bulk wave generating device is configured to generate a bulk wave, the bulk wave enables the bubble to resonate for generating a flow field; and the surface acoustic wave generating device is configured to generate a surface acoustic wave and control a position of at least one particle in the solution.

In various embodiments methods are provided for delivering an agent of interest (e.g., protein, antibody, nucleic acid) into cells. In certain embodiments the method comprises contacting the cells with anisotropic magnetic particles in the presence of the agent; and applying a substantially uniform magnetic field to said magnetic particles where movement of said particles induced by said magnetic field introduces transient openings into said cell facilitating entry of said agent of interest into said cells.

Improved biolistic bombardment is observable when using a modification to a double-barrel device. One such modification is a custom attachment that can be retrofit to a standard helium-based gene gun. The custom attachment facilitates DNA and protein delivery. The attachment confines the flow of particles in a barrel similar to the choke on a shotgun barrel.

Delivering a payload across a plasma membrane of a cell includes providing a population of cells and contacting the population of cells with a volume of an aqueous solution. The aqueous solution includes the payload and alcohol content greater than 5 percent concentration. The volume of the aqueous solution may be a function of exposed surface area of the population of cells, or may be a function of a number of cells in the population of cells. Related compositions, apparatus, systems, techniques, and articles are also described.

The invention relates to bacterial artificial chromosomes (BAC) comprising retroviral nucleic acid sequences encoding: gag and pol proteins, and an env protein or a functional substitute thereof, wherein each of the retroviral nucleic acid sequences are arranged as individual expression constructs within the BAC. The invention also relates to uses and methods of transient transfection using said BAC.

The invention relates to bacterial artificial chromosomes (BAC) comprising retroviral nucleic acid sequences encoding: gag and pol proteins, and an env protein or a functional substitute thereof, wherein each of the retroviral nucleic acid sequences are arranged as individual expression constructs within the BAC. The invention also relates to uses and methods of transient transfection using said BAC.