Cell expansion

Patent metadata

Field	Value
Publication number	US-11965175-B2
Application number	US-201716302951-A
Country	US
Kind code	B2
Filing date	May 25, 2017
Priority date	May 25, 2016
Publication date	Apr 23, 2024
Grant date	Apr 23, 2024

Abstract

Embodiments are described that relate to methods and systems for growing cells in a hollow fiber bioreactor. In embodiments, the cells may be exposed to a number of growth factors including a combination of recombinant growth factors. In other embodiments, the cells may be grown in co-culture with other cells, e.g., hMSC's. In embodiments, the cells may include CD34+ cells.

First claim

What is claimed is:

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.

2. The method of claim 1 , wherein the first plurality of cells is derived from cord blood.

3. The method of claim 1 , wherein the human mesenchymal stem cells are bone marrow derived.

4. The method of claim 3 , further comprising:
administering the second plurality of expanded cells to a patient to reconstitute hematopoiesis in the patient.

5. The method of claim 1 , further comprising:
flushing the hollow fiber bioreactor with a washing fluid prior to introducing the first plurality of cells.

6. The method of claim 1 , wherein expanding the first plurality of cells comprises controlling an environment of the hollow fiber bioreactor.

7. The method of claim 6 , wherein controlling the environment of the hollow fiber bioreactor comprises:
monitoring at least one parameter within the hollow fiber bioreactor; and
adjusting the at least one parameter to keep the at least one parameter within an acceptable range.

8. The method of claim 7 , wherein adjusting the at least one parameter comprises controlling a flow rate of the growth media flowing through the extracapillary side of the hollow fiber bioreactor.

Assignees

• Terumo Bct Inc

Inventors

• Jones Mark E

Classifications

• A61K35/15 Primary

  Cells of the myeloid line, e.g. granulocytes, basophils, eosinophils, neutrophils, leucocytes, monocytes, macrophages or mast cells; Myeloid precursor cells; Antigen-presenting cells, e.g. dendritic cells (presenting a specific antigen A61K39/00; therapeutic combinations of antibodies, or fragments thereof, and blood-derived cells A61K39/00) · CPC title

• C12N5/0068 Primary

  General culture methods using substrates (for specific animal cell type C12N5/06) · CPC title

• C12M1/10

  rotatably mounted · CPC title

• C12M3/00

  Tissue, human, animal or plant cell, or virus culture apparatus · CPC title

• C12M25/10

  Hollow fibers or tubes (hollow fiber modules in general B01D63/02) · CPC title