An object of the invention is to provide an electroporation method for treating vesicles with exogenous material for insertion of the exogenous material into the vesicles which includes the steps of: a. retaining a suspension of the vesicles and the exogenous material in a treatment volume in a chamber which includes electrodes, wherein the chamber has a geometric factor (cm.sup.−1) defined by the quotient of the electrode gap squared (cm.sup.2) divided by the chamber volume (cm.sup.3), wherein the geometric factor is less than or equal to 0.1 cm.sup.−1, wherein the suspension of the vesicles and the exogenous material is in a medium which is adjusted such that the medium has conductivity in a range spanning 50 microSiemens/cm to 500 microSiemens/cm, wherein the suspension is enclosed in the chamber during treatment, and b. treating the suspension enclosed in the chamber with one or more pulsed electric fields. With the method, the treatment volume of the suspension is scalable, and the time of treatment of the vesicles in the chamber is substantially uniform.

An object of the invention is to provide an electroporation method for treating vesicles with exogenous material for insertion of the exogenous material into the vesicles which includes the steps of: a. retaining a suspension of the vesicles and the exogenous material in a treatment volume in a chamber which includes electrodes, wherein the chamber has a geometric factor (cm.sup.−1) defined by the quotient of the electrode gap squared (cm.sup.2) divided by the chamber volume (cm.sup.3), wherein the geometric factor is less than or equal to 0.1 cm.sup.−1, wherein the suspension of the vesicles and the exogenous material is in a medium which is adjusted such that the medium has conductivity in a range spanning 50 microSiemens/cm to 500 microSiemens/cm, wherein the suspension is enclosed in the chamber during treatment, and b. treating the suspension enclosed in the chamber with one or more pulsed electric fields. With the method, the treatment volume of the suspension is scalable, and the time of treatment of the vesicles in the chamber is substantially uniform.

The present invention relates to a process for producing a composition comprising an aqueous medium and, disposed in the aqueous medium, a first volume of a first hydrogel, which process comprises: (i) providing a composition comprising a first hydrophobic medium and, disposed in the first hydrophobic medium, a first volume of a first hydrogel; (ii) disposing a volume of an aqueous composition comprising a hydrogel compound around the first volume of the first hydrogel; (iii) allowing the aqueous composition comprising the hydrogel compound to form a gel and thereby forming a hydrogel object, which hydrogel object comprises the first volume of the first hydrogel and a second volume of a second hydrogel, which second volume of the second hydrogel is disposed around the first volume of the first hydrogel; and (iv) transferring the hydrogel object from the first hydrophobic medium to an aqueous medium and thereby producing the composition comprising the aqueous medium and, disposed in the aqueous medium, the first volume of the first hydrogel. The invention further provides a hydrogel object, which hydrogel object comprises a first volume of a first hydrogel and a second volume of a second hydrogel, which second volume of the second hydrogel is disposed around the first volume of the first hydrogel.

The present invention relates to a process for producing a composition comprising an aqueous medium and, disposed in the aqueous medium, a first volume of a first hydrogel, which process comprises: (i) providing a composition comprising a first hydrophobic medium and, disposed in the first hydrophobic medium, a first volume of a first hydrogel; (ii) disposing a volume of an aqueous composition comprising a hydrogel compound around the first volume of the first hydrogel; (iii) allowing the aqueous composition comprising the hydrogel compound to form a gel and thereby forming a hydrogel object, which hydrogel object comprises the first volume of the first hydrogel and a second volume of a second hydrogel, which second volume of the second hydrogel is disposed around the first volume of the first hydrogel; and (iv) transferring the hydrogel object from the first hydrophobic medium to an aqueous medium and thereby producing the composition comprising the aqueous medium and, disposed in the aqueous medium, the first volume of the first hydrogel. The invention further provides a hydrogel object, which hydrogel object comprises a first volume of a first hydrogel and a second volume of a second hydrogel, which second volume of the second hydrogel is disposed around the first volume of the first hydrogel.

A cell culture device (1) includes: a culture vessel (10 ) that accommodates a cell suspension containing cells and a culture medium; and a stirring device (30) that is provided at a bottom part of the culture vessel (10) to stir the cell suspension accommodated in the culture vessel (10). The stirring device (30) has a shaft part (32), and a rotating part (33) rotatable with the shaft part (32) as a rotation axis. The rotating part (33) has a hole part (36) into which the shaft part (32) is inserted, and a gap (37) of 100 μm or more is secured between an interior wall demarcating the hole part (36) and the shaft part (32). A closing part (38) that closes an end part of the gap (37) in an axial direction of the rotation axis is provided.

A cell culture device (1) includes: a culture vessel (10 ) that accommodates a cell suspension containing cells and a culture medium; and a stirring device (30) that is provided at a bottom part of the culture vessel (10) to stir the cell suspension accommodated in the culture vessel (10). The stirring device (30) has a shaft part (32), and a rotating part (33) rotatable with the shaft part (32) as a rotation axis. The rotating part (33) has a hole part (36) into which the shaft part (32) is inserted, and a gap (37) of 100 μm or more is secured between an interior wall demarcating the hole part (36) and the shaft part (32). A closing part (38) that closes an end part of the gap (37) in an axial direction of the rotation axis is provided.

The present disclosure provides a membrane separation method of a cell suspension which can appropriately separate cells from debris, and a cell culture device. That is, membrane separation processing of the cell suspension is performed using a filtration membrane which includes an inlet-side opening formed on one surface and an outlet-side opening, which is formed on the other surface and communicates with the inlet-side opening, and in which the inlet-side opening and the outlet-side opening are disposed at positions deviated in a direction parallel to the surfaces of the membrane.

The present disclosure provides a membrane separation method of a cell suspension which can appropriately separate cells from debris, and a cell culture device. That is, membrane separation processing of the cell suspension is performed using a filtration membrane which includes an inlet-side opening formed on one surface and an outlet-side opening, which is formed on the other surface and communicates with the inlet-side opening, and in which the inlet-side opening and the outlet-side opening are disposed at positions deviated in a direction parallel to the surfaces of the membrane.

Described herein are FasL-engineered biomaterials, as well as methods of making and using such FasL-engineered biomaterials, such as for immunomodulation, such as for inducing immunosuppression and specific immune tolerance, such as for preventing or reducing the risks of rejection of cellular or tissue grafts and/or the treatment of autoimmune disorders such as Type I diabetes. In specific embodiments, the FasL-engineered biomaterials are biotinylated microgels bound to SA-FasL.

Provided is an observation device including: a stereo image-acquisition optical system that acquires images of cells floating in a culture fluid inside a culture vessel; and an analyzer that calculates a cell density of the cells on the basis of the images acquired by the stereo image-acquisition optical system, wherein the analyzer identifies a three-dimensional position of each of the cells included in the images and calculates the cell density on the basis of the number of cells present within a predetermined three-dimensional region.