A spheroid cell culture article including: a frame having a chamber including: an opaque side wall surface; a top aperture; a gas-permeable, transparent bottom; and optionally a chamber annex surface and second bottom, and at least a portion of the transparent bottom includes at least one concave arcuate surface, is disclosed. Methods of making and using the article are also disclosed.

This invention provides a cell culture substrate comprising on its surface a fluorine-containing polymer that enables three-dimensional tissue culture. The cell culture substrate of the invention has a surface at least a part of which is composed of a resin composition comprising a fluorine-containing polymer having one or more fluorine atoms in a repeating unit and exhibits the oxygen gas permeability of 219 cm.sup.3 (STP)/(m.sup.2.Math.24 h.Math.atm) or higher. Three-dimensional tissue can be formed via cell culture with the use of the cell culture substrate of the invention.

Cell culture systems and methods provide improved immunotherapeutic product manufacturing with greater scalability, flexibility, and automation. Cell culture systems are configured with interchangeable cartridges, allowing versatility and scalability. Systems are configured to have multiple connected cell culture chambers, which allows parallel processing of different types of cells. Gas-impermeable cell culture chambers and methods for generating cells in closed systems prevent contamination and user error. Methods for recycling cell culture medium provide additional efficiencies.

Methods of production of edible filamentous fungal biomat formulations are provided as standalone protein sources and/or protein ingredients in foodstuffs as well as a one-time use or repeated use self-contained biomat reactor comprising a container with at least one compartment and placed within the compartment(s), a feedstock, a fungal inoculum, a gas-permeable membrane, and optionally a liquid nutrient medium.

This invention relates to methods and devices that improve cell culture efficiency. They include the use of gas permeable culture compartments that reduce the use of space while maintaining uniform culture conditions, and are more suitable for automated liquid handling. They include the integration of gas permeable materials into the traditional multiple shelf format to resolve the problem of non-uniform culture conditions. They include culture devices that use surfaces comprised of gas permeable, plasma charged silicone and can integrate traditional attachment surfaces, such as those comprised of traditional tissue culture treated polystyrene. They include culture devices that integrate gas permeable, liquid permeable membranes. A variety of benefits accrue, including more optimal culture conditions during scale up and more efficient use of inventory space, incubator space, and disposal space. Furthermore, labor and contamination risk are reduced.

A culture device and a culture method by which a substance to be cultured can be three-dimensionally cultured, and the substance thus cultured can be removed as an integrated product with multiple tubes; and a cultured organ produced by the culture method. The culture device comprises a sealed container (2) which contains the substance to be cultured (A) and is disassembled after the culture is completed; multiple ducts (3, 4, 5) arranged in the sealed container (2) and have micropore formed on the outer peripheral surface; a culture medium-feeding device (6, 7) for feeding/circulating the culture medium (B) to at least one duct (3 or 5); an excretory device (8) connected to at least one duct (4) for excreting the waste product (C) permeating into the duct (4) through the micropore of the duct (4) from the substance to be cultured to the outside of the sealed container.

A cell culture container includes a container main body comprising a gas-permeable plastic film, and an injection/discharge port. A peripheral portion of the container main body is sealed, and the container main body has a bulging shape on a top surface side of the container main body. A plurality of concave portions serving as cell culture parts are provided on a bottom surface of the container main body.

An apparatus is provided for the production of biomass or a bioproduct, the apparatus comprising: at least one elongate bioreactor, the bioreactor comprised of at least one outer membrane layer that defines a substantially tubular compartment that is capable of being filled with a liquid or gel, wherein the membrane layer is comprised of a material that is permeable to gas transfer across the membrane layer. A chamber is provided comprising walls that define and enclose a gaseous atmosphere within. At least a part of the bioreactor is located inside the chamber. A control system controls the composition of the atmosphere within the chamber and gas transfer occurs across the membrane layer of the bioreactor between the tubular compartment and the atmosphere comprised within the chamber. Methods of using the apparatus in order to manufacture biomass are also provided.

A polymer or other substrate optimized for growing cells is described, which takes the form of a micro-thin bag with gas permeable sides. Sides of the bag can be held at a fixed distance from one another with a multitude of tiny micropillars or other spacers extending between them, keeping the bag at a predetermined thickness and preventing the bag from collapsing and the sides from sticking together. In other embodiments, the sides may be held apart by gas pressure alone. A 0.01 μm to 1000 μm parylene or other biocompatible coating over the bag outsides controls the permeability of the bag material and provides a bio-safe area for cell growth. An alternate configuration uses open-cell foam with skins coated with a biocompatible coating. Tubes going into multiple bags can be connected to a manifold that delivers gaseous oxygen or removes carbon dioxide and other waste gases. Multiple bags can be stacked together tightly, with o-ring spacers in between, and housed within a vessel to form a high-surface area, ultra-compact cell growing system. For cells growing on the bags, liquid nutrients can be fed by way of the tube spacers, and oxygen and waste gases permeated through the bag sides and transported within the bags.

A culture container for culturing epithelial cells, includes an upper container, a lid member configured to airtightly fit with an opening portion of the upper container, and a lower container configured to accommodate the upper container and a cell culture medium, in which at least a part of an area of the upper container in contact with the cell culture medium is formed of a membrane that is permeable to at least a part of components of the cell culture medium and impermeable to a cell, and a material of the lid member has an oxygen permeability coefficient of 1.0×10.sup.−6 cm.sup.3 cm/(cm.sup.2.Math.sec.Math.atm) or less.