Expanding cells in a bioreactor

Inventors

NANKERVIS, Brian J. • HILL, Domicinda M. • Jones, Mark E.

Assignees

Terumo BCT Inc

Abstract

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.

Core Innovation

A method of expanding cells in a cell expansion system is described by loading a first volume of fluid comprising a plurality of cells into a first fluid flow path of the cell expansion system, where a bioreactor is fluidly associated with the first fluid flow path. The method includes loading media into a portion of the first fluid flow path to position the first volume of fluid in a first portion of the bioreactor, exposing the plurality of cells to an activator, and feeding and expanding the plurality of cells during a first time period.

During a second time period, feeding is performed according to a second process that includes pumping media at a first flow rate into a first port of the bioreactor and concurrently pumping media at a second flow rate into a second port of the bioreactor. The direction of the second flow rate into the bioreactor is opposite the direction of the first flow rate into the bioreactor, and the plurality of cells is expanded during the second time period under these concurrent, oppositely directed pumping conditions.

After expanding during the first time period and after expanding during the second time period, the plurality of cells is circulated at a first circulation rate during a third time period to reduce a number of cells in a first cell cluster and a second cell cluster. The disclosed system architecture includes intracapillary and extracapillary spaces with IC/EC loops, inlet ports and outlet ports, together with an oxygenation module, sensors, and a rocking device configured for closed, automated operation.

Claims Coverage

One independent claim is provided. The claim covers a multi-stage cell expansion method with activator exposure, first and second feeding and expansion time periods, concurrent oppositely directed media pumping into first and second bioreactor ports, and circulation steps to reduce cell clusters; dependent claims further narrow the cell population and include an explicit numeric concentration constraint for the loaded cell-containing fluid.

The core coverage is the combination of activator exposure with two distinct feeding and expansion time periods, where the second feeding process uses concurrent media pumping into first and second bioreactor ports with opposite flow directions, and where circulation at a first circulation rate is applied after expansion to reduce cells in first and second cell clusters. Dependent claims narrow the cell type to non-adherent cells and/or T cells, including regulatory T cells, and add an explicit concentration bound for the loaded first volume of cell-containing fluid.

Stated Advantages

Documented Applications