Cell expansion
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.
1. A method of expanding cells in a cell expansion system, the method comprising:
loading a first volume of a first fluid into the cell expansion system, wherein the first fluid comprises a plurality of cells, and wherein the cell expansion system comprises a cell growth chamber;
based on the first volume of the first fluid, determining a second volume of a second fluid to load into a portion of a first fluid circulation path to position the first volume of the first fluid in a first portion of the cell growth chamber, wherein the second fluid comprises media, and wherein the first portion of the cell growth chamber comprises a central portion of the cell growth chamber;
based on the determining the second volume of the second fluid, loading the second volume of the second fluid into the cell expansion system to position the first volume of the first fluid in the central portion of the cell growth chamber;
loading a third volume of a third fluid for feeding the plurality of cells; and
expanding the plurality of cells.
2. The method of claim 1 , wherein the cell expansion system comprises a second fluid circulation path.
3. The method of claim 1 , wherein the second volume of the second fluid in the first fluid circulation path flows through an intracapillary space of the cell growth chamber.
4. The method of claim 3 , wherein a fourth fluid in the second fluid circulation path flows through an extracapillary space of the cell growth chamber.
5. The method of claim 1 , wherein the first volume of the first fluid is loaded without activating an intracapillary circulation pump.
6. The method of claim 1 , wherein the first volume of the first fluid is the same as the second volume of the second fluid.
7. The method of claim 1 , wherein the first volume of the first fluid is different from the second volume of the second fluid.
8. The method of claim 1 , wherein the cell growth chamber comprises a hollow fiber membrane.
9. The method of claim 1 , further comprising:
stimulating the plurality of cells with an activator to activate the plurality of cells.
10. The method of claim 9 , wherein the activator comprises a soluble activator.
11. The method of claim 9 , wherein the activator comprises one or more beads.
12. The method of claim 1 , wherein the plurality of cells comprises non-adherent cells.
13. The method of claim 12 , wherein the plurality of cells comprises T cells.
14. The method of claim 13 , wherein the plurality of cells comprises a subpopulation of T cells.
15. The method of claim 14 , wherein the subpopulation of T cells comprises regulatory T cells (Tregs).
16. The method of claim 1 , wherein the loading the first volume of the first fluid comprises loading the first volume of the first fluid without activating a circulation pump.
17. The method of claim 1 , further comprising:
determining to reposition the first volume of the first fluid in the cell growth chamber; and
based on the determining to reposition the first volume of the first fluid, loading a fourth volume of a fourth fluid into the first fluid circulation path.
18. The method of claim 1 , wherein the second volume of the second fluid includes a first amount of the second fluid in a first portion of the first fluid circulation path.
19. The method of claim 18 , wherein the second volume of the second fluid includes a second amount of the second fluid in a second portion of the first fluid circulation path.
20. The method of claim 19 , wherein the second portion of the first fluid circulation path comprises an intracapillary space between an inlet port of the cell growth chamber and the central portion of the cell growth chamber.