Cell capture and expansion
AssigneeTerumo BCT, Inc.
InventorsMark E. Jones, Dalip Sethi, Dennis J. Hlavinka, Thomas J. Felt
Life of this patent
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Abstract
Implementations are described that relate to methods and systems for growing cells in a hollow fiber bioreactor. In implementations, the cells may be exposed to a number of growth factors including a combination of recombinant growth factors. In other implementations, the cells may be grown in co-culture with other cells, e.g., hMSC's. In implementations, the cells may include CD34+ cells. A coated membrane includes a membrane having a first coating configured to promote cellular adhesion to the membrane and a second coating that includes a soluble protein moiety.
Claims26
A method of expanding cells, comprising:
- applying a coating to an interior lumen of a plurality of hollow fibers of a hollow fiber bioreactor, the plurality of hollow fibers being semi-permeable, the coating including fibronectin configured to adhere to the interior lumen and stromal cell-derived factor 1 (SDF-1) configured to both adhere to the fibronectin and retain CD34+ hematopoietic stem cells thereto;
- introducing a first plurality of cells into the plurality of hollow fibers of the hollow fiber bioreactor, the first plurality of cells including CD34+ hematopoietic stem cells derived from at least one of cord blood, bone marrow, and peripheral blood;
- maintaining and concentrating the first plurality of cells within the plurality of hollow fibers by directing fluid into opposite ends of the hollow fiber bioreactor simultaneously;
- exposing the first plurality of cells in the hollow fibers to growth conditions;
- and
- expanding at least a portion of the first plurality of cells in the hollow fibers of the bioreactor to generate a second plurality of expanded target cells including CD34+ hematopoietic stem cells that are expanded at least 50-fold, by introducing into the plurality of hollow fibers, and into contact with the first plurality of cells, the following:
- soluble growth supporting cytokines including at least one of SCF, TPO, Flt-3L, IL-3, and IL-6;
- an aryl receptor antagonist;
- and
- glial cell-derived neurotrophic factor (GDNF).
Dependencies are inferred from the claim text.