Patent ID: 11965175
Assignee: TERUMO BCT, INC.
Field: Pharmaceuticals (Chemistry)
Classification: CPC C  A | IPC A  C

Claim 0:
1. A method of expanding cells, the method comprising: coating with fibronectin an intracapillary side of a plurality of hollow fibers extending through a bioreactor;
introducing human mesenchymal stem cells (hMSCs) into the plurality of hollow fibers coated with fibronectin by activating a pump to pump the hMSCs into the plurality of hollow fibers, stopping the pump to allow a first portion of the hMSCs to attach to a first area of an inside of the plurality of hollow fibers coated with fibronectin, rotating the bioreactor 180° from an initial start position, reactivating the pump to circulate a second portion of the hMSCs within the hollow fibers, and stopping the pump to allow the second portion of the hMSCs to attach to a second area of the inside of the hollow fibers coated with fibronectin that is opposite to the first area;
culturing the hMSCs in a first media;
introducing a first plurality of cells comprising CD34+ cells into the plurality of hollow fibers and rotating the bioreactor to form a co-culture of CD34+ cells and hMSCs,
wherein the first plurality of cells is added to the hollow fiber bioreactor without purification;
exposing the co-culture of CD34+ cells and hMSCs to a second media, which is a growth media including a soluble combination of recombinant FMS-like tyrosine kinase 3 ligand (rhFlt-3L), recombinant human stem cell factor (rhSCF), recombinant thrombopoietin (rhTPO), and recombinant human glial cell-derived neurotrophic factor (rhGDNF);
expanding at least a portion of the first plurality of cells in the plurality of hollow fibers of the bioreactor to generate a second plurality of expanded cells by circulating nutrients including glucose the growth media through an extracapillary side of the hollow fiber bioreactor; and
removing the second plurality of expanded cells from the bioreactor.