Abstract: A formulation for a culture medium that is specifically designed to supplement a culture of differentiating hematopoietic stem cells (HSCs) with the factors necessary for high density manufacture of red blood cells (RBCs) while only requiring partial medium exchanges on a periodic basis. Further, the present disclosure identifies a factor, iron (III) citrate, also referred to as ferric citrate, that can supplant more commonly utilized sources of biological iron in culture medium for a more stable, more cost-effective medium formulation for superior vertical scalability.

Abstract: A method includes directing fluid flow from a bioreactor to an external path of a re-circulation loop, separating cells from the medium and returning the cells to the bioreactor thereby leaving spent medium, depleting the spent medium of biological waste products, replenishing the spent medium with beneficial growth factors to thereby provide replenished medium, and directing the replenished medium back to the bioreactor to thereby complete the re-circulation loop such that recycling of the medium occurs. The replenishing of the spent medium with beneficial growth factors may occur within a medium collection reservoir. The method may further include monitoring the spent medium within the medium collection reservoir including temperature of the spent medium within the medium collection reservoir and pH of the spent medium within the medium collection reservoir.

Abstract: A system includes a bioreactor having an inlet and an outlet, a cell-media recirculation loop between the outlet of the bioreactor and the inlet of the bioreactor, and a mature cell separation unit positioned along the cell-media recirculation loop wherein the mature cell separation unit is configured for performing separation of mature cells from immature cells and direction of the immature cells back to the inlet of the bioreactor. The system may further include a storage unit for storing the mature cells. The mature cell separation unit may be further configured to direct the mature cells to a mature cell storage unit.