Abstract: Systems and methods for treating media in large-scale bioreactors can include a method of using a large-scale bioreactor system. The method can include (a) mixing a mixture of at least cells, and media in a bioreactor, and (b) removing a portion of the media through an output port of the bioreactor. The method can also include (c) retaining the cells within the bioreactor using a screen covering the port. The method can also include (d) receiving a second media, which may in some examples include the portion of the first media removed from the bioreactor, into a media preparation container external to the bioreactor, (e) treating the second media within the media preparation container to produce oxygenated media within a threshold pH range, and (f) introducing the oxygenated media into the bioreactor using a return port of the bioreactor.

Abstract: Systems and methods for complete medium exchange using two bioreactors and at least one external separation and retention device designed for therapeutic cells grown as aggregates, on the surface of microcarriers, or as single cells, for scalable expansion and/or directed differentiation.

Abstract: Material transfer devices and related systems and methods are disclosed. In accordance with an example, a material transfer device includes a housing including a first housing portion and a second housing portion and a screen. The first housing portion includes a first inlet port, a first outlet port, and a first transfer opening. The second housing portion has a second inlet port, a second outlet port, and a second transfer opening. The first transfer opening is disposed adjacent to and in communication with the second transfer opening. The screen is disposed between the housing portions adjacent to the first and second transfer openings.

Abstract: Systems and methods for scalable manufacturing of therapeutic cells in bioreactors are disclosed. Fluid dynamic considerations for scale in accordance with an implementation include a method of production of therapeutic cells grown on microcarriers or as cell aggregates in a suspension-based bioreactor includes depositing a suspension comprising cells suspended in a volume of culture fluid into a bioreactor and setting an agitation rate of a mixer disposed in the bioreactor. The method includes actuating the mixer at the set agitation rate to mix the suspension in the bioreactor. The suspension includes a plurality of turbulent eddies generated by the mixer. A magnitude of an energy dissipation rate (EDR) of at least approximately 60% of the turbulent eddies can be less than approximately 0.0015 m2/s3.

Abstract: A vessel having a mixer that ensures a homogeneous cell distribution in dispensed quantities. The vessel has a mixer therein for stirring contents of the vessel and an orifice in a lower wall to which a cell dispenser is attached. The cell dispenser dispenses quantities of suspended cells having a homogeneous cell distribution. The vessel may be a closed system with no option or instructions for removing a cap or other access port, whereby after manufacturing the vessel is shipped to a customer and the interior remains closed to the exterior environment and fluids are transferred through tubes.

Abstract: Macrocarriers with flat surface areas which can be easily suspended in liquid using low power input will improve cell attachment and growth in bioreactors. Such macrocarriers can be consistently manufactured to exact size specifications, which can significantly reduce the risk of contamination by small size particulates. The macrocarriers may be flat discs or squares with central bumps on each face to prevent stacking. The macrocarriers may also have outward curved wings from two or four sides to facilitate distribution in moving solution.

Abstract: Methods of repeated aggregate dissociation and reformation of pluripotent stem cells (PSCs) within the same bioreactor until a desired final cell number is achieved. A preferred step-wise process for controlled growth of PSCs and aggregate size using periodic dissociation with a dissociation medium which contains either proteolytic enzymes or chemical reagents, mechanical agitation, or a combination of these methods.

Abstract: A vessel having a mixer that ensures a homogeneous cell distribution in dispensed quantities. The vessel has a mixer therein for stirring contents of the vessel and an orifice in a lower wall to which a cell dispenser is attached. The cell dispenser dispenses quantities of suspended cells having a homogeneous cell distribution.

Abstract: The bioreactor may include a single-use, rigid-sided bioreactor vessel containing a fluid to be mixed and a vertical mixing wheel. A perfusion dip tube with a screen filter incorporated on the end is secured to the vessel from the top lid and partially submerged in the fluid, preferably into close proximity with an outer circumference of the vertical mixing wheel. The screen filter of appropriate mesh size allows spent cell culture medium to be withdrawn from the bioreactor vessel while retaining cell aggregates or microcarriers on which cells are attached and growing in the vessel. Alternating flow of fluid out from and into the dip tube enables removal of spent medium and unclogging of the dip tube filter.

Abstract: A system and method for compartmentalizing micro-carriers in a bioreactor includes a container configured to store micro-carriers and a vessel configured to contain a fluid so as to prevent the micro-carriers from entering the vessel during sterilization, shipping, storage, or other pre-use handling of the system. One or more addition lines connect the container and the vessel such that the vessel is in fluid communication with the vessel. The one or more addition lines are contactable by at least one blocking element configured to reversibly block fluid flow between the container and the vessel. The container, addition lines, and vessel are configured to allow the micro-carriers to be injected into the vessel at any point during a cell culture run. The vessel may also include a rotatable wheel, a harvest port configured to allow for the removal of the micro-carriers from the vessel, and a media removal port comprising a retention screen for removing spent medium.

Abstract: A system and method for compartmentalizing micro-carriers in a bioreactor includes a container configured to store micro-carriers and a vessel configured to contain a fluid so as to prevent the micro-carriers from entering the vessel during sterilization, shipping, storage, or other pre-use handling of the system. One or more addition lines connect the container and the vessel such that the vessel is in fluid communication with the vessel. The one or more addition lines are contactable by at least one blocking element configured to reversibly block fluid flow between the container and the vessel. The container, addition lines, and vessel are configured to allow the micro-carriers to be injected into the vessel at any point during a cell culture run. The vessel may also include a rotatable wheel, a harvest port configured to allow for the removal of the micro-carriers from the vessel, and a media removal port comprising a retention screen for removing spent medium.

Abstract: A vessel having a mixer that ensures a homogeneous cell distribution in dispensed quantities. The vessel has a mixer therein for stirring contents of the vessel and an orifice in a lower wall to which a cell dispenser is attached. The cell dispenser dispenses quantities of suspended cells having a homogeneous cell distribution.

Abstract: A system and method for compartmentalizing micro-carriers in a bioreactor includes a container configured to store micro-carriers and a vessel configured to contain a fluid so as to prevent the micro-carriers from entering the vessel during sterilization, shipping, storage, or other pre-use handling of the system. One or more addition lines connect the container and the vessel such that the vessel is in fluid communication with the vessel. The one or more addition lines are contactable by at least one blocking element configured to reversibly block fluid flow between the container and the vessel. The container, addition lines, and vessel are configured to allow the micro-carriers to be injected into the vessel at any point during a cell culture run. The vessel may also include a rotatable wheel, a harvest port configured to allow for the removal of the micro-carriers from the vessel, and a media removal port comprising a retention screen for removing spent medium.

Abstract: A system and method for compartmentalizing micro-carriers in a bioreactor includes a container configured to store micro-carriers and a vessel configured to contain a fluid so as to prevent the micro-carriers from entering the vessel during sterilization, shipping, storage, or other pre-use handling of the system. One or more addition lines connect the container and the vessel such that the vessel is in fluid communication with the vessel. The one or more addition lines are contactable by at least one blocking element configured to reversibly block fluid flow between the container and the vessel. The container, addition lines, and vessel are configured to allow the micro-carriers to be injected into the vessel at any point during a cell culture run. The vessel may also include a rotatable wheel, a harvest port configured to allow for the removal of the micro-carriers from the vessel, and a media removal port comprising a retention screen for removing spent medium.

Abstract: The present invention relates generally to a method for increasing the yield of plasmid DNA production. The method includes the steps of selecting a highly productive clonal subtype of a strain of E. coli, including but not limited to the DH5 strain, harboring a DNA plasmid and cultivating said clonal subtype with fed-batch fermentation in a chemically-defined medium. The plasmid DNA production process described herein can generate record quantities of plasmid DNA when said highly productive clonal subtypes are cultivated on an industrial scale. The disclosed method can be used for the production of pharmaceutical grade DNA for use in polynucleotide vaccination and gene therapy treatment regimens.

Abstract: An aseptic sampling system includes substantially sealed sampling lines connecting one or more sample container to a vessel containing the fluid to be sampled. A flow control system includes valves and/or clamps placed strategically along the sampling lines and filtered vent lines are provided at strategic locations relative to a reversible fluid pump in communication with the sampling lines. The clamps are selectively actuated, the vent lines are selectively opened or closed, and the pump is operated in a forward or reverse direction to permit a volume of sample fluid to be drawn from the vessel and into a sample container and to permit fluid to be purged from the sampling lines either into the sample container or back into the vessel. Thus, sampling and purging can be accomplished without opening the system to potentially contaminating agents and without wasting or having to properly dispose the fluid.

Abstract: The present invention relates generally to a method for increasing the yield of plasmid DNA production. The method includes the steps of selecting a highly productive clonal subtype of a strain of E. coli, including but not limited to the DH5 strain, harboring a DNA plasmid and cultivating said clonal subtype with fed-batch fermentation in a chemically-defined medium. The plasmid DNA production process described herein can generate record quantities of plasmid DNA when said highly productive clonal subtypes are cultivated on an industrial scale. The disclosed method can be used for the production of pharmaceutical grade DNA for use in polynucleotide vaccination and gene therapy treatment regimens.

Abstract: The present invention relates generally to a method for increasing the yield of plasmid DNA production. The method includes the steps of selecting a highly productive clonal subtype of a strain of E. coli, including but not limited to the DH5 strain, harboring a DNA plasmid and cultivating said clonal subtype with fed-batch fermentation in a chemically-defined medium. The plasmid DNA production process described herein can generate record quantities of plasmid DNA when said highly productive clonal subtypes are cultivated on an industrial scale. The disclosed method can be used for the production of pharmaceutical grade DNA for use in polynucleotide vaccination and gene therapy treatment regimens.

Abstract: An aseptic sampling system includes substantially sealed sampling lines connecting one or more sample container to a vessel containing the fluid to be sampled. A flow control system includes valves and/or clamps placed strategically along the sampling lines and filtered vent lines are provided at strategic locations relative to a reversible fluid pump in communication with the sampling lines. The clamps are selectively actuated, the vent lines are selectively opened or closed, and the pump is operated in a forward or reverse direction to permit a volume of sample fluid to be drawn from the vessel and into a sample container and to permit fluid to be purged from the sampling lines either into the sample container or back into the vessel. Thus, sampling and purging can be accomplished without opening the system to potentially contaminating agents and without wasting or having to properly dispose the fluid.

Abstract: A bioreactor includes a fluid containment vessel and a rotating mixing element including radial inflow elements configured to direction fluid radially inwardly of the mixing element and axial out flow elements configured to redirect the radial inward flow in an axial direction. Both the vessel and the mixing element are made from plastic and the vessel comprises a flexible bag supported by a rigid housing. In one embodiment, the mixing element is pneumatically driven by the buoyancy of gas introduced into the vessel and trapped within gas entrapment cups formed on the periphery of the mixing element. As the volumetric capacity of the bioreactor increases, the ratio of the diameter of the mixing element to the width of the vessel decreases, and a gap defined between the bottom of the mixing element and the bottom of the vessel increase.