This invention relates to compositions of matter, methods, modules and instruments for automated mammalian cell growth and mammalian cell transduction followed by nucleic acid-guided nuclease editing in live mammalian cells.

Compositions of matter, methods, modules, and automated instruments may relate to synthesizing a library including an editing cassette including a different gRNA and donor DNA pair, amplifying the editing cassette in a partition separate from other editing cassettes in the library, adding nuclease to the partition, and adding lipofectamine to the editing cassette and nuclease to form a lipofectamine/nucleic acid/nuclease complex. A microcarrier coated in extracellular matrix or a cell adhesion molecule coating may be added to the lipofectamine/nucleic acid/nuclease complex. Cell growth material, the microcarrier, and mammalian cells may be transferred to a growth module in an automated closed cell editing instrument via a liquid handling system. The mammalian cells may be allowed to seed on the microcarrier. Conditions may be provided for the mammalian cells to take-up and be edited by a payload associated with the lipofectamine/nucleic acid/nuclease complex. The mammalian cells may be detached from the microcarrier.

The present disclosure relates to a microcarrier for cell culture comprising: polystyrene-based particles containing at least one or more of hydrocarbon oil having 12 or more carbon atoms, or pores derived therefrom, a method for producing the same, and a cell culture composition using the same.

The devices, methods, and compositions disclosed herein accomplish robust cell encapsulation in polymer microparticles using a vertically oriented microfluidic device. A hydrophilic polymer precursor solution is flowed into a first inlet channel, which extends inward from an upper surface of the device housing. A hydrophobic fluid is flowed into a second inlet channel, which extends inward from a lower surface of the device housing. The two inlet channels meet at a junction, and an outlet channel extends away from the two inlet channels. When the two inwardly flowing streams meet at the junction, the polymer precursor solution disperses into the hydrophobic fluid. The dispersed precursor droplets are photopolymerized into microparticles as they travel through the outlet channel. The resulting microparticles are highly uniform, and are larger than conventionally formed microparticles. Cells of varying types can be encapsulated with high viability and spatial uniformity.

Described herein are apparata and methods for growing cells in a manner that mimics the native three-dimensional environment. Cell cultures grown in the apparatus can be screened for inhibition by specific chemotherapeutics or other drugs.

The present invention relates to a method and system for aseptic material transfer from a storage container to a bioreactor. More precisely the invention relates to a method and system for aseptically transferring dry material, such as microcarriers for cell cultivation, to a bioreactor, comprising transferring microcarriers from a first container housing said microcarriers to a bioreactor for cell cultivation via r transfer tubing connecting the first container to the bioreactor, wherein the transfer is accomplished with pressurized air or gas supplied to the container.

The present disclosure provides a microfluidic device for simulating a blood-brain barrier and a blood-brain barrier simulation system including the same, and the microfluidic device includes: a first channel; a second channel which is adjacently connected to the first channel through one or more microholes and configured to culture neural stem cells; and a chamber which is connected to both ends of the first channel and contains a culture medium.

A filter assembly for separating microcarriers from a fluid medium includes a collapsible container bounding a sterile compartment adapted to hold a fluid. An inlet port is attached to the container through which fluid flows into the compartment. An outlet port is attached to the container through which fluid flows out of the compartment. A filter is disposed within the compartment, the filter dividing the compartment into an inlet chamber that is fluidly coupled with the inlet port and an outlet chamber that is fluidly coupled with the outlet port, the filter allowing a medium to pass therethrough but preventing microcarriers disposed in the medium from passing therethrough.

The subject invention concerns materials and methods for expansion of stem cells, such as mesenchymal stem cells (MSC), that improve translational success of the cells in the treatment of various conditions. The subject invention utilizes cell self-aggregation as a non-genetic means to enhance their therapeutic potency in a microcarrier bioreactor. In one embodiment of the method cells are cultured in a container or vessel in the presence of thermally responsive microcarriers (TRMs) wherein cells adhere to the surface of the TRMs. After a period of time the cell culture temperature is reduced so that the cells detach from the TRMs. The detached cells are allowed to form 3D aggregates. The 3D aggregates can be collected and treated to dissociate the cells. Dissociated cells can then be used for transplantation in methods of treatment or for in vitro characterization and study.

A process and apparatus for the enzymatic degradation of a cellulosic substrate is disclosed. The process comprises agitating a composition with milling particles, wherein the milling particles are or comprise a lignocellulosic material and wherein the composition comprises: a. the cellulosic substrate; b. a cellulase enzyme; and c. a liquid medium.