Embodiments described herein generally provide for the expansion of cells in a cell expansion system using an active promotion of a coating agent(s) to a cell growth surface. A coating agent may be applied to a surface, such as the cell growth surface of a hollow fiber, by controlling the movement of a fluid in which a coating agent is suspended. Using ultrafiltration, the fluid may be pushed through the pores of a hollow fiber from a first side, e.g., an intracapillary (IC) side, of the hollow fiber to a second side, e.g., an extracapillary (EC) side, while the coating agent is actively promoted to the surface of the hollow fiber. In so doing, the coating agent may be hydrostatically deposited onto a wall, e.g., inner wall, of the hollow fiber.

A cell culture system having a cell culture vessel, a composition controlling fluid storage vessel, a culture fluid composition controlling means having an inlet and an outlet for a cell culture fluid, an inlet-connected fluid feeding circuit from the cell culture vessel to the inlet of the culture fluid composition controlling means, an outlet-connected fluid feeding circuit from the cell culture vessel to the outlet of the culture fluid composition controlling means, a means which perfuses the cell culture fluid from the inlet-connected fluid feeding circuit to the outlet-connected fluid feeding circuit through the culture fluid composition controlling means, and a means which controls the amount of fluid in the cell culture vessel, in which compositions of the cell culture fluid in the cell culture vessel and compositions of the composition controlling fluid in the composition controlling fluid storage vessel can be controlled in a continuous manner.

This disclosure describes methods and apparatuses for creating or using a fibrous cell substrate that is formed in a mold using temperature and wettability gradients and that upon which cells can be grown. The disclosed method can include utilizing a temperature gradient and a wettability gradient within a mold to form a porous cell substrate. In particular, the temperature and wettability gradients control nucleation and growth of crystals within the cell substrate solution to form parallel layers of cell substrate. The crystals can be sublimated by freeze drying and the porous cell substrate is seeded. More specifically, the disclosed method includes using pressure and an increased cell mixture viscosity to seed cells deep within the cell substrate. This disclosure also describes an apparatus for forming the structured porous cell substrate.

Embodiments described herein generally provide for expanding cells in a cell expansion system. The cells may be grown in a bioreactor, and the cells may be activated by an activator (e.g., a soluble activator complex). Nutrient and gas exchange capabilities of a closed, automated cell expansion system may allow cells to be seeded at reduced cell seeding densities, for example. Parameters of the cell growth environment may be manipulated to load the cells into a particular position in the bioreactor for the efficient exchange of nutrients and gases. System parameters may be adjusted to shear any cell colonies that may form during the expansion phase. Metabolic concentrations may be controlled to improve cell growth and viability. Cell residence in the bioreactor may be controlled. In embodiments, the cells may include T cells. In further embodiments, the cells may include T cell subpopulations, including regulatory T cells (Tregs), for example.

Described are embodiments for expanding cells in a bioreactor. In one embodiment, methods are provided that distribute cells throughout the bioreactor and attach cells to specific portions of a bioreactor to improve the expansion of the cells in the bioreactor. Embodiments may be implemented on a cell expansion system configured to load, distribute, attach and expand cells.

A waste treatment, pyrolysis and gasification and concerns an apparatus for syngas bio-methanation include a unit for pyrolysis/gasification receiving organic material, the unit for pyrolysis/gasification generating syngas, comprising at least one membrane reactor inside a liquid bath comprising at least one bacteria population, the membrane reactor comprising at least one hollow fiber in contact with the liquid bath, around which a biofilm is formed and into which the syngas from the unit for pyrolysis/gasification flows, so as to convert the syngas into methane. A method for bio-methanation of syngas comprising a step of providing syngas from a unit for pyrolysis/gasification to a membrane reactor inside a liquid bath comprising at least one suitable bacteria population, the membrane reactor comprising at least one hollow fiber in contact with the liquid bath, around which a biofilm is formed and into which the output syngas of the unit for pyrolysis flows, so as to convert the syngas into methane.

This invention is directed to methods of directly reseeding harvested adherent cells grown in a hollow fiber bioreactor. Also disclosed is a novel harvest media for use in directly reseeding adherent cells into a hollow fiber bioreactor.

One or more embodiments are described directed to a method and system for loading and distributing cells in a bioreactor of a cell expansion system. Accordingly, embodiments include methods and systems that may provide for adding a plurality of cells to a fluid within a bioreactor of the cell expansion system. A first percentage of the plurality of cells is allowed to settle in the bioreactor and a second percentage of the plurality of cells is allowed to settle outside of the bioreactor. The first percentage of cells is then expanded in the bioreactor. The second percentage of cells is wasted.

The invention is to provide a cell culture substrate excellent in cellular adhesiveness regardless of a shape or design thereof. Provided is a cell culture substrate comprising a coating layer on at least one side of a polymer substrate, wherein the coating layer includes at least one member selected from the group consisting of a copolymer (a) comprising 40% by mole or more and 90% by mole or less of a structural unit (a-1) derived from alkoxyalkyl (meth)acrylate of following Formula (1) and 10% by mole or more and 60% by mole or less of a structural unit (a-2) derived from trialkyl aminoalkyl (meth)acrylate of following Formula (2), a copolymer (b) comprising 85% by mole or more and 95% by mole or less of a structural unit (b-1) derived from alkoxyalkyl (meth)acrylate of following Formula (1) and 5% by mole or more and 15% by mole or less of a structural unit (b-2) derived from carboxyalkyl (meth)acrylate of following Formula (3), and a copolymer (c) comprising 70% by mole or more and 90% by mole or less of a structural unit (c-1) derived from alkoxyalkyl (meth)acrylate of following Formula (1) and 10% by mole or more and 30% by mole or less of a structural unit (c-2) derived from hydroxyalkyl (meth)acrylate of following Formula (4).

This invention is to provide a means capable of obtaining excellent cell proliferation activity without depending on a thickness of a coating layer in a technique of coating a cell culture substrate (cell culture vessel) using a polymer. Provided is a cell culture substrate comprising a coating layer on at least one surface of a polymer substrate, wherein the coating layer includes a copolymer comprising more than 40% by mole and less than 100% by mole of a structural unit (1) derived from carboxyalkyl (meth)acrylate represented by Formula (1) and more than 0% by mole and less than 60% by mole of a structural unit (2) derived from ethylenically unsaturated monomer having a hydroxyl group (the total of the structural unit (1) and the structural unit (2) is 100% by mole).