Abstract: A method for in vivo RNA or protein expression is provided. The method includes introducing a double-stranded concatemeric DNA into a eukaryotic cell to generate a desired RNA or protein. The double-stranded concatemeric DNA includes a plurality of tandem repeat sequences, wherein each of the plurality of tandem repeat sequences comprises an expression sequence. The double-stranded concatemeric DNA comprises one or more phosphorothioated nucleotides, wherein a ratio of phosphorothioated nucleotides to total nucleotides in the double-stranded concatemeric DNA is at least 1:1600. A eukaryotic cell comprising an exogeneous, double-stranded concatemeric DNA comprising the plurality of tandem repeat sequences is also provided.

Abstract: Provided herein are novel methods for expansion and passaging of cell aggregates comprising stem cells and/or differentiated cells and comprising the use of closed systems in stirred tank bioreactors. The methods of the invention permit closed system serial passage expansion of pluripotent stem cells and/or progeny thereof with associated pluripotency markers and differentiation potential.

Abstract: A carrier for expansion of pluripotent stem cells is provided, wherein the carrier comprises a substrate comprising one or more outer surfaces, wherein the one or more outer surfaces are modified with gas plasma treatment, and one or more structured indentations on one or more of the outer surfaces. The carrier has a length at least about 0.2 mm, a width at least about 0.2 mm, and a height in a range from about 0.05 mm to 1.2 mm and each of the structured indentations has a major axis in a range from about 0.1 mm to 0.5 mm, a minor axis in a range from about 0.1 mm to 0.5 mm and a depth in a range from about 0.025 mm to about 0.5 mm. A method of making the carrier, and culturing stromal cells using the same carrier are also provided.

Abstract: Provided herein are novel methods for expansion and passaging of cell aggregates comprising stem cells and/or differentiated cells and comprising the use of closed systems on rocking platform bioreactors. The methods of the invention permit closed system serial passage expansion of pluripotent stem cells and/or progeny thereof with associated pluripotency markers and differentiation potential.

Abstract: Described are multi-functional beads and methods to capture rare cells directly from low-volume biological samples and perform both functional and genomic assays from those cells. This is accomplished using a multifunctional capture bead that allows co-localization of both the single cell capture element and the molecular assay components. When combined with a digital microfluidic platform this enables encoding and/or barcoding of specific single cells.

Abstract: A carrier for expansion of pluripotent stem cells is provided, wherein the carrier comprises a substrate comprising one or more outer surfaces, wherein the one or more outer surfaces are modified with gas plasma treatment, and one or more structured indentations on one or more of the outer surfaces. The carrier has a length at least about 0.2 mm, a width at least about 0.2 mm, and a height in a range from about 0.05 mm to 1.2 mm and each of the structured indentations has a major axis in a range from about 0.1 mm to 0.5 mm, a minor axis in a range from about 0.1 mm to 0.5 mm and a depth in a range from about 0.025 mm to about 0.5 mm. A method of making the carrier, and culturing stromal cells using the same carrier are also provided.

Abstract: A carrier for expansion of induced pluripotent stem cells is provided, wherein the carrier comprises a substrate comprising one or more outer surfaces, wherein the one or more outer surfaces are modified with gas plasma treatment, and one or more structured indentations on one or more of the outer surfaces. The carrier has a length at least about 0.2 mm, a width at least about 0.2 mm, and a height in a range from about 0.05 mm to 1.2 mm and each of the structured indentations has a major axis in a range from about 0.1 mm to 0.5 mm, a minor axis in a range from about 0.1 mm to 0.5 mm and a depth in a range from about 0.025 mm to about 0.5 mm. A method of making the carrier, and culturing induced pluripotent stem cells using the same carrier are also provided.

Abstract: A carrier for expansion of pluripotent stem cells is provided, wherein the carrier comprises a substrate comprising one or more outer surfaces, wherein the one or more outer surfaces are modified with gas plasma treatment, and one or more structured indentations on one or more of the outer surfaces. The carrier has a length at least about 0.2 mm, a width at least about 0.2 mm, and a height in a range from about 0.05 mm to 1.2 mm and each of the structured indentations has a major axis in a range from about 0.1 mm to 0.5 mm, a minor axis in a range from about 0.1 mm to 0.5 mm and a depth in a range from about 0.025 mm to about 0.5 mm. A method of making the carrier, and culturing stromal cells using the same carrier are also provided.

Abstract: Provided herein are novel methods for expansion and passaging of cell aggregates comprising stem cells and/or differentiated cells and comprising the use of closed systems on rocking platform bioreactors. The methods of the invention permit closed system serial passage expansion of pluripotent stem cells and/or progeny thereof with associated pluripotency markers and differentiation potential.

Abstract: Provided herein are novel methods for expansion and passaging of cell aggregates comprising stem cells and/or differentiated cells and comprising the use of closed systems in stirred tank bioreactors. The methods of the invention permit closed system serial passage expansion of pluripotent stem cells and/or progeny thereof with associated pluripotency markers and differentiation potential.

Abstract: A method of eliciting an immune response in an organism comprising: providing an unprocessed rolling circle amplification (RCA) product; and administering an effective amount of the unprocessed RCA product to the organism to elicit the immune response, wherein the unprocessed RCA product is prepared from a circular nucleic acid template comprising at least one promoter sequence, and at least one target sequence. A vaccine comprising unprocessed RCA product is also provided and methods for making the same.

Abstract: A carrier for expansion of induced pluripotent stem cells is provided, wherein the carrier comprises a substrate comprising one or more outer surfaces, wherein the one or more outer surfaces are modified with gas plasma treatment, and one or more structured indentations on one or more of the outer surfaces. The carrier has a length at least about 0.2 mm, a width at least about 0.2 mm, and a height in a range from about 0.05 mm to 1.2 mm and each of the structured indentations has a major axis in a range from about 0.1 mm to 0.5 mm, a minor axis in a range from about 0.1 mm to 0.5 mm and a depth in a range from about 0.025 mm to about 0.5 mm. A method of making the carrier, and culturing induced pluripotent stem cells using the same carrier are also provided.

Abstract: A method of delivering exogenous molecules, comprising: providing a plurality of cells having a cell membrane; adding a plurality of exogenous molecules to the cells; exposing the cells to a defocused infrared (IR) light to permeabilize the cell membrane of the cells; and delivering the exogenous molecules to the cells through the permeablized cell membrane, wherein an intensity of the IR light at the optical focus is at least greater than or equal to an order of 104 W/cm2.

Abstract: An embodiment of a system is provided herein, wherein the system allows for the analysis and selection of numerous experimental conditions to optimize transfection efficiency and cell viability. The system is used for magnetic particle based nucleic acid delivery by optimizing various parameters. The system comprises a control module; an incubation module for incubating magnetic nanoparticle and nucleic acid; a transfection module and an analysis module.

Abstract: A system for delivering exogenous molecules comprises a support for containing cells and exogenous molecules; an infra-red (IR) light source that generates an IR optical beam with an average power density at least greater than 105 W/cm2; one or more optical elements; an imaging system to image the cells in a field of view; a processor that generates a signal for localization of cells in the field of view; a light pattern shaper for temporal focusing of optical beam to generate wide field illumination on the cells to permeabilise the cell membrane for delivering the exogenous molecules; and a controller that switches optical beam from wide field illumination to a focused illumination. The processor is operatively coupled to the imaging system and the light pattern shaper and transmits the signal for the localization of cells to ensure the temporal focusing of the optical beam on the cells.

Abstract: A method of delivering exogenous molecules, comprising: providing a plurality of cells having a cell membrane; adding a plurality of exogenous molecules to the cells; exposing the cells to a defocused infrared (IR) light to permeabilize the cell membrane of the cells; and delivering the exogenous molecules to the cells through the permeablized cell membrane, wherein an intensity of the IR light at the optical focus is at least greater than or equal to an order of 104 W/cm2.

Abstract: The invention relates generally to a process of analyzing and visualizing the expression of biomarkers in individual cells wherein the cells are examined to develop patterns of expression by using a grouping algorithm, and a system to perform and display the analysis.

Abstract: A bushing assembly for a turbine vane includes a bushing, housing, and a flexible conductor. The bushing includes a bore for receiving a trunnion of the vane and a first conductor extending from the bushing. The first conductor is adapted to be connected to a heater element disposed on the vane. The housing is positioned over the bushing, and the bushing is rotatable relative to the housing. The housing includes a second conductor extending from the housing for connection to an electrical power source. The flexible conductor is electrically connected between the first conductor and the second conductor, such that an electric current is transmitted from the second conductor to the first conductor to provide an electric current to the heater element.

Abstract: A bushing assembly for a turbine vane includes a bushing and a housing. The bushing includes a bore for receiving a trunnion of the vane, a first electrical contact carried on an outer surface thereof, and a first conductor extending from the first electrical contact. The first electrical contact is adapted to be connected to a heater element disposed on the vane. The housing is positioned over the bushing and rotatable relative to the bushing. The housing includes a second electrical contact disposed in contact with the first electrical contact and a second conductor extending from the second electrical contact for connection to an electrical power source.

Abstract: A method of eliciting an immune response in an organism comprising: providing an unprocessed rolling circle amplification (RCA) product; and administering an effective amount of the unprocessed RCA product to the organism to elicit the immune response, wherein the unprocessed RCA product is prepared from a circular nucleic acid template comprising at least one promoter sequence, and at least one target sequence. A vaccine comprising unprocessed RCA product is also provided and methods for making the same.