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), helper, naïve, memory, or effector, for example.

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), helper, naïve, memory, or effector, for example.

The present application relates to a bioreactor system for culturing cells, especially cells without cell walls; the bioreactor system includes: a container containing a hollow cylinder with a diameter of D1 and a height of H1, and a hollow circular truncated cone with an upper diameter of D2, a lower diameter of D3, and a height of H2, where the hollow cylinder is connected to a top surface of the hollow circular truncated cone, and D1=D2; an oscillator, configured to cause the container to make an eccentric motion according to a certain eccentricity and rotational speed; a ventilation device, configured to introduce an oxygen-containing gas from an upper portion of the container to the inside of the container, and a culture solution filled in the container, of which a top surface is exposed to the oxygen-containing gas; where the oscillator is configured to maintain the eccentric motion of the container, such that a ratio of the total liquid surface area to the volume (S/V) of the culture solution when in a steady state of motion is 5.65 or more, the turbulence kinetic energy is 2.73E−03 m.sup.2/s.sup.2 or more, and the flow field shear rate is 20.27/s or less, where the total liquid surface area is the sum of the contact area between the culture solution and the reactor wall surface and the contact area between the top surface and the gas.

The present application relates to a bioreactor system for culturing cells, especially cells without cell walls; the bioreactor system includes: a container containing a hollow cylinder with a diameter of D1 and a height of H1, and a hollow circular truncated cone with an upper diameter of D2, a lower diameter of D3, and a height of H2, where the hollow cylinder is connected to a top surface of the hollow circular truncated cone, and D1=D2; an oscillator, configured to cause the container to make an eccentric motion according to a certain eccentricity and rotational speed; a ventilation device, configured to introduce an oxygen-containing gas from an upper portion of the container to the inside of the container, and a culture solution filled in the container, of which a top surface is exposed to the oxygen-containing gas; where the oscillator is configured to maintain the eccentric motion of the container, such that a ratio of the total liquid surface area to the volume (S/V) of the culture solution when in a steady state of motion is 5.65 or more, the turbulence kinetic energy is 2.73E−03 m.sup.2/s.sup.2 or more, and the flow field shear rate is 20.27/s or less, where the total liquid surface area is the sum of the contact area between the culture solution and the reactor wall surface and the contact area between the top surface and the gas.

The present invention provides a culture vessel and a measuring device for measuring a cell sheet thickness nondestructively during culturing. A culture vessel having a lid 4 has a float 8 installed in the interior and the float is structured so as to be uplifted by a culture medium in the culture vessel. Further, a cell sheet thickness measuring device having a sensor head 10 having a light-emitting element 10A and a light-receiving element 10B to detect a positional variation of the float in the vertical direction and a controller 11 to process a signal of the sensor head is configured.

The present invention provides a culture vessel and a measuring device for measuring a cell sheet thickness nondestructively during culturing. A culture vessel having a lid 4 has a float 8 installed in the interior and the float is structured so as to be uplifted by a culture medium in the culture vessel. Further, a cell sheet thickness measuring device having a sensor head 10 having a light-emitting element 10A and a light-receiving element 10B to detect a positional variation of the float in the vertical direction and a controller 11 to process a signal of the sensor head is configured.

A method for producing cell aggregates includes culturing cells while suspending the cells in a liquid culture medium comprising a lysophospholipid. A composition includes a lysophospholipid, wherein the composition is a liquid culture medium or a composition added to a liquid culture medium.

A method for producing cell aggregates includes culturing cells while suspending the cells in a liquid culture medium comprising a lysophospholipid. A composition includes a lysophospholipid, wherein the composition is a liquid culture medium or a composition added to a liquid culture medium.

A method for dissociating cell aggregates in an agitated reactor. The method comprises providing a cell culture comprising cell aggregates in the agitated reactor, contacting the cell aggregates with a dissociation reagent, generating a dissociation force in the agitated reactor and exposing the contacted cell aggregates to the generated dissociation force under conditions sufficient to dissociate the contacted cell aggregates. The method may be used in a process for passaging cells and/or generating dissociated differentiated cells from stem and/or progenitor cells.

A method for dissociating cell aggregates in an agitated reactor. The method comprises providing a cell culture comprising cell aggregates in the agitated reactor, contacting the cell aggregates with a dissociation reagent, generating a dissociation force in the agitated reactor and exposing the contacted cell aggregates to the generated dissociation force under conditions sufficient to dissociate the contacted cell aggregates. The method may be used in a process for passaging cells and/or generating dissociated differentiated cells from stem and/or progenitor cells.