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.

A multi-layer microfluidic bioreactor is used in methods for culturing cells. A grooved semipermeable substrate can capture and grow cells using perfusion culture methods. Microfluidic geometry directs flow over grooves for cell expansion and along grooves for cell harvesting. The bioreactor and methods can be optimized for high density and automated processing for growth of cells.

The disclosure relates generally to a cell culture apparatus and a cell culture method. One aspect of the disclosure is an oxygen-permeable bag comprising one or more polymer films defining a boundary of an interior compartment of the bag, each of the one or more polymer films comprising an inner layer adjacent the interior compartment of the bag, the inner layer comprising a fluoropolymer, and adhered to the inner layer, an outer layer comprising polymer; a first port formed in an exterior surface of the bag and in fluid communication with the interior compartment; a second port formed in an exterior surface of the bag and in fluid communication with the interior compartment; and contained in the interior compartment, a plurality of microcarriers.

An improved method of culturing cells for cell therapy applications that includes growing desired cells in the presence of antigen-presenting cells and/or feeder cells and with medium volume to surface area ratio of up to 1 ml/cm.sup.2 if the growth surface is not comprised of gas permeable material and up to 2 ml/cm.sup.2 if the growth surface is comprised of gas permeable material. The desired cells are at a surface density of less than 0.5×10.sup.6 cells/cm.sup.2 at the onset of a production cycle, and the surface density of the desired cells plus the surface density of the antigen presenting cells and/or feeder cells are at least about 1.25×10.sup.5 cells/cm.sup.2.

A cell culture bag includes a body formed of a composite film including a first fluoropolymer and a second fluoropolymer. The body defines a cell culture compartment configured to hold a cell culture. The first fluoropolymer has a first thickness and the second fluoropolymer at least partially impregnates the first thickness of the first fluoropolymer. The composite film has a second thickness from 0.01 mm to 0.059 mm and a tensile strength in one direction of 10,000 psi to 92,000 psi. The composite film also has O.sub.2 flux of 2,000 cm.sup.3/M.sup.2/atm/day to 20,000 cm.sup.3/M.sup.2/atm/day and a total transmittance from 70% to 100%. A cell culture assembly includes a plurality of the cell culture bags connected in series. A cell culture container includes a cell culture compartment and a composite film connected to the cell culture compartment, the composite film including a first fluoropolymer and a second fluoropolymer.

A cell activation reactor and a cell activation method are provided. The cell activation reactor includes a body, a rotating part, an upper cover, a microporous film, and multiple baffles. The body has an accommodating space, which is suitable for accommodating multiple cells and multiple magnetic beads. The rotating part is disposed in the accommodating space and includes multiple impellers. The microporous film is disposed in the accommodating space and covers multiple holes of the accommodating space. The baffles are disposed in the body. When the rotating part is driven to rotate, the interaction between the baffles and the impellers separates the cells and the magnetic beads.

A multilayered cell culture apparatus for the culturing of cells is disclosed. The cell culture apparatus is defined as an integral structure having a plurality of cell culture chambers in combination with tracheal space(s). The body of the apparatus has imparted therein gas permeable membranes in combination with tracheal spaces that will allow the free flow of gases between the cell culture chambers and the external environment. The flask body also includes an aperture that will allow access to the cell growth chambers by means of a needle or cannula. The size of the apparatus, and location of an optional neck and cap section, allows for its manipulation by standard automated assay equipment, further making the apparatus ideal for high throughput applications.

An in vitro expansion process for rapid expansion of antigen specific T cells, such as allogeneic antigen specific cells comprising the steps culturing in a gas permeable vessel a population of PBMCs (such as allogeneic PBMCs) in the presence of antigen, for example a peptide or peptide mix relevant to a target antigen(s), in the presence of an exogenous cytokine characterized in that the expansion to provide the desired population of T cells is 14 days or less, for example 9, 10, 11 or 12 days, such as 10 days. The disclosure also extends to T cell populations generated by and obtained from the method and the use of same in therapy.

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.

The present set of embodiments relate to a system, method, and apparatus for separating cells from a suspension using a gravity driven cell separation device. The apparatus may include a chamber that is separated into a gas compartment and a liquid compartment allowing for transfer of gas through a membrane into the liquid compartment where the cell suspension flows during operation. The embodiments are designed to maximize the gravitational separation effect on the suspension while mitigating the loss of surface area through which gas transfer can occur through a membrane to provide nutrients to cells in the suspension by tilting the cell separation device on two or more axes.