Simultaneous stimulation and concentration of cells

• US 6,797,514 B2
• Filed: 09/20/2001
• Issued: 09/28/2004
• Est. Priority Date: 02/24/2000
• Status: Expired due to Term

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Xcyte Therapies, Inc. (Cyclacel Pharmaceuticals Incorporated)
• Inventors: Bonyhadi, Mark, Kalamasz, Dale, Craig, Stewart, Hardwick, Alan, Saund, Narinder, Law, Che, Berenson, Ronald, McMillen, David
• Primary Examiner(s): Lankford, Jr., Leon B.
• Application Number:
  US09/960,264
• Publication Number:
  US 20020119568A1
• Time in Patent Office: 1,104 Days
• Field of Search: 435/325, 435/375, 435/372.3, 436/526
• US Class Current: 435/325
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61K 2039/57 characterised by the type of response, e.g. Th1, Th2
• CPC Class Codes: A61L 27/3804 characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
• CPC Class Codes: A61L 27/3895 using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions
• CPC Class Codes: C07K 16/2803 against the immunoglobulin superfamily
• CPC Class Codes: C07K 16/2806 against CD2
• CPC Class Codes: C07K 16/2809 against the T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 16/2812 against CD4
• CPC Class Codes: C07K 16/2815 against CD8
• CPC Class Codes: C07K 16/2818 against CD28 or CD152
• CPC Class Codes: C07K 16/2821 against ICAM molecules, e.g. CD50, CD54, CD102
• CPC Class Codes: C07K 16/2827 against B7 molecules, e.g. CD80, CD86
• CPC Class Codes: C07K 16/2833 against MHC-molecules, e.g. HLA-molecules
• CPC Class Codes: C07K 16/2845 against integrin beta2-subunit-containing molecules, e.g. CD11, CD18
• CPC Class Codes: C07K 16/2854 against selectins, e.g. CD62
• CPC Class Codes: C07K 16/2866 against receptors for cytokines, lymphokines, interferons
• CPC Class Codes: C07K 16/2875 against the NGF/TNF superfamily, e.g. CD70, CD95L, CD153, CD154 against NGF C07K16/22, against TNF C07K16/241
• CPC Class Codes: C07K 16/2878 against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
• CPC Class Codes: C07K 16/289 against CD45
• CPC Class Codes: C07K 16/2896 against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C12N 2500/32 : Amino acids
• CPC Class Codes: C12N 2501/23 : Interleukins [IL]
• CPC Class Codes: C12N 2501/51 : B7 molecules, e.g. CD80, CD86, CD28 (ligand), CD152 (ligand)
• CPC Class Codes: C12N 2501/515 : CD3, T-cell receptor complex
• CPC Class Codes: C12N 2501/53 : CD2
• CPC Class Codes: C12N 2533/12 : Glass
• CPC Class Codes: C12N 2533/14 : Ceramic
• CPC Class Codes: C12N 2533/18 : Calcium salts, e.g. apatite, Mineral components from bones, teeth, shells
• CPC Class Codes: C12N 2533/30 : Synthetic polymers
• CPC Class Codes: C12N 2533/40 : Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers
• CPC Class Codes: C12N 2533/54 : Collagen; Gelatin
• CPC Class Codes: C12N 2533/70 : Polysaccharides
• CPC Class Codes: C12N 2533/72 : Chitin, chitosan
• CPC Class Codes: C12N 2533/90 : Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
• CPC Class Codes: C12N 5/0635 : B lymphocytes
• CPC Class Codes: C12N 5/0636 : T lymphocytes
• CPC Class Codes: C12N 5/0647 : Haematopoietic stem cells; Uncommitted or multipotent progenitors

Abstract

The present invention relates generally to methods for stimulating cells, and more particularly, to a novel method to concentrate and stimulate cells that maximizes stimulation and/or proliferation of such cells. In the various embodiments, cells are stimulated and concentrated with a surface yielding enhanced proliferation, cell signal transduction, and/or cell surface moiety aggregation. In certain aspects methods for stimulating a population of cells such as T-cells, by simultaneous concentration and cell surface moiety ligation are provided by contacting the population of cells with a surface, that has attached thereto one or more agents that ligate a cell surface moiety and applying a force that predominantly drives cell concentration and cell surface moiety ligation, thereby inducing cell stimulation, cell surface moiety aggregation, and/or receptor signaling enhancement. Also provided are methods for producing phenotypically tailored cells, including T-cells for the use in diagnostics, drug discovery, and the treatment of a variety of indications, including cancer, viral infection, and immune related disorders. Compositions of cells having specific phenotypic properties produced by these processes are further provided.

Claims

1. A device adapted for the magnetic attraction of particles in a cell culture comprising a cell culture vessel adapted for the culture of human cells and a magnet or a magnetizable element, wherein said magnet or magnetizable element is positioned within a wall of said culture vessel or external to said culture vessel so as to allow a magnetic field emanating therefrom to pass through said culture vessel.

2. The device of claim 1, wherein said magnet or magnetizable element is an electromagnet.

3. The device of claim 1, wherein said human cells are lymphocytes.

4. The device of claim 1, wherein said culture vessel is a flask or a bag.

5. A method for inducing aggregation of cells or cell surface markers comprising:

(a) providing a population of cells having a target cell surface molecule; and

(b) providing a device, wherein said device is adapted for the magnetic attraction of particles in a cell culture and wherein said device comprises

a cell culture vessel adapted for the culture of human cells, and

a magnet or a magnetizable element, wherein said magnet or magnetizable element is positioned so as to allow a magnetic field emanating therefrom to pass through said culture vessel; and

wherein said cells are contained within said culture vessel;

said culture vessel contains a solid surface having attached thereto a ligand for at least one target cell surface molecule; and

(c) applying a magnetic field through said magnet or magnetizable element of said device,

thereby inducing aggregation of said cells or cell surface markers.

6. The method of claim 5, wherein said aggregation induces target cell signal transduction.

7. The method of claim 5, wherein said solid surface is selected from the group consisting of a plate, a bag, a dish, a rod, a pellet, a fiber, a microsphere, and a bead.

8. The method of claim 7, wherein said solid surface is a bead or a microsphere.

9. The method of claim 7, wherein said solid surface is paramagnetic.

10. The method of claim 5, wherein said cell population comprises lymphocytes.

11. The method of claim 5, wherein said ligand is selected from the group consisting of an antibody, a natural ligand, and a synthetic ligand.

12. The method of claim 5, wherein said ligand comprises an antibody, a peptide, a polypeptide, a growth factor, a cytokine, or a chemokine.

13. The method of claim 5, wherein said receptor binding leads to downregulation or suppression of a cellular event.

14. The method of claim 5, wherein said receptor binding leads to upregulation or activation of a cellular event.