The present invention relates to cell culture in bioreactors, such as flexible cellbag bioreactors. More specifically the present invention relates to methods for simplifying the production of clinically relevant cell products for use in cell therapy.

Provided is a cell culturing method in which physical stress is reduced and oxygen can be supplied. The invention relates to a cell culturing method including; arranging cells in a dispersed state in a culture medium which is a plastic fluid; and introducing air bubbles into said culture medium.

A method has been developed to enhance the efficacy of cancer vaccines by activating the immune system against a greater variety of antigens expressed in the tumor cells. In this modification, the vaccine is created against not only the more mature cancer cells, but also cancer stem cells (CSCs), that act as tumor propagating cells, and can also be made against as the more mature progeny of the CSCs that are normally present within the malignant tumors in numbers which are too low to effectively manufacture a vaccine against their antigens, but which are responsible for recurrence of the malignant tumor. These include pluripotent and stem cells induced from cells in a tumor biopsy by exposure to stress inducing agents that cause the cells to almost die, thereby causing cells to de-differentiate. The method greatly increases the variety of the tumor antigens at which the vaccine is targeted.

Described herein are methods and compositions for use in expanding alveolar epithelial cells. The methods may include the use of three-dimensional substrates and improved techniques for expansion of the cells. The improved composition for culturing alveolar epithelial cells may include at least one or more of the following: a TGF- pathway inhibitor; a Wnt pathway activator; a ROCK inhibitor; an epidermal growth factor (EGF); a keratinocyte growth factor (KGF); and a fetal bovine serum.

Cell-culture platforms are described that can impart cyclic strain on a biologically-relevant culture substrate. Accordingly, systems and methods of the invention can provide a more accurate model of physiological strain and forces than earlier technologies. Systems of the invention enable high-throughput experimentation in standard culturing equipment, while utilizing a 3D physiologically-relevant environment.

The present invention provides a biomolecular composite which is a ghost cell which has been emptied of all or substantially all of the cytosolic contents of the cell including the nucleus and nuclear contents, preferably further having been fused with exogenous amphipathic molecules, wherein said composite has a hydrodynamic diameter of less than 6 m. The biomolecular composite may contain a cargo molecule and be capable of targeting a cell or tissue. Also provided are methods of preparing ghost cells, methods of preparing biomolecular composites and methods for their use.

A method for manufacturing a cell mass adherently cultured on a cell culture substrate, in which the cell culture substrate comprises two regions (A) and (B), and the method comprising steps (1) to (3): (A) an island-shaped region having a cell proliferation property and an area of 0.001 to 1 mm.sup.2; and (B) a region adjacent to the region (A) and having no cell proliferation properties, (1) a step of preparing a cell suspension comprising a mesenchymal cell and an ectoderm-derived cell; (2) a step of bringing the cell suspension into contact with the cell culture substrate, and randomly adhering the two types of cells to the region (A), and in which, on the contact surface between the cell suspension and the cell culture substrate, a total number of cells of the two types of cells is 100 to 100,000 cells/cm.sup.2; and (3) a step of culturing the cell adhered to the region (A) and forming a cell mass comprising the two types of cells.

One aspect of the invention provides an apparatus for isolating one or more subcellular components including a cell disruption reservoir that generates at least one of a phase change, a thermal change, a physical contact force, an ultrasonic frequency, an osmotic change, a pressure change, a photothermal pulse, a magnetic field, an electromagnetic field, an electric field, and an electrical pulse through the reservoir and a separation instrument configured to specifically isolate the subcellular components based on one or more parameters selected from at least one of density, charge/pH, dielectric polarization, magnetic attraction, spectral dispersion, spectral refraction, spectral diffraction, hydrophobicity, hydrophilicity, structure (presence or absence of a structural feature), function (migration), affinity or binding, and pressure.

Provided is a method for efficiently harvesting cells cultured at a large scale from a three-dimensional porous scaffold. This method includes: a culturing step in which cells are cultured on a three-dimensional porous scaffold sealed in a culture vessel; an enzymatic treatment step in which an enzyme solution containing trypsin-EDTA, caseinase, and/or collagenase is added to the culture vessel to conduct an enzymatic treatment on the cultured cells in the culture vessel; and a vibration treatment step in which a vibration treatment is conducted, i.e., a base that makes a circular motion within a horizontal plane and a bottom surface of the culture vessel are made to come into surface contact with one another so as to impart a vertical-direction vibration to the cultured cells that have undergone the enzymatic treatment.

Disclosed herein are methods for generating satellite cells and compositions including satellite cells.