Method for obtaining monocytes or NK cells

• US 10,113,148 B2
• Filed: 07/10/2013
• Issued: 10/30/2018
• Est. Priority Date: 08/02/2012
• Status: Active Grant

• Current Assignee: Hiroaki Kawasaki, Hiroyuki Abe
• Original Assignee: Hiroaki Kawasaki, Hiroyuki Abe
• Inventors: Abe, Hiroyuki, Kawasaki, Hiroaki
• Primary Examiner(s): Wehbe, Anne Marie S
• Application Number:
  US14/414,377
• Publication Number:
  US 20150197727A1
• Time in Patent Office: 1,938 Days
• Field of Search: 435347, 435372, 53038873
• CPC Class Codes: A61K 2039/5158 Antigen-pulsed cells, e.g. T-cells
• CPC Class Codes: C07K 16/2896 against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C12N 2500/90 Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2501/02 Compounds of the arachidonic acid pathway, e.g. prostaglandins, leukotrienes
• CPC Class Codes: C12N 2501/22 Colony stimulating factors (G-CSF, GM-CSF)
• CPC Class Codes: C12N 2501/2301 Interleukin-1 (IL-1)
• CPC Class Codes: C12N 2501/2302 Interleukin-2 (IL-2)
• CPC Class Codes: C12N 2501/2304 Interleukin-4 (IL-4)
• CPC Class Codes: C12N 2501/2306 Interleukin-6 (IL-6)
• CPC Class Codes: C12N 2501/2312 Interleukin-12 (IL-12)
• CPC Class Codes: C12N 2501/2315 Interleukin-15 (IL-15)
• CPC Class Codes: C12N 2501/25 Tumour necrosing factors [TNF]
• CPC Class Codes: C12N 2501/26 Flt-3 ligand (CD135L, flk-2 ligand)
• CPC Class Codes: C12N 2501/58 Adhesion molecules, e.g. ICAM, VCAM, CD18 (ligand), CD11 (ligand), CD49 (ligand)
• CPC Class Codes: C12N 2501/599 with CD designations not provided for elsewhere
• CPC Class Codes: C12N 2506/115 from monocytes, from macrophages
• CPC Class Codes: C12N 5/0639 Dendritic cells, e.g. Langherhans cells in the epidermis
• CPC Class Codes: C12N 5/0645 Macrophages, e.g. Kuepfer cells in the liver; Monocytes
• CPC Class Codes: C12N 5/0646 Natural killers cells [NK], NKT cells

Abstract

The present invention relates to a method by which cells usable for an immune cell therapy are separated from peripheral blood and grown. The present invention makes it possible to provide immune system cells which are large enough in number to be used in the immune cell therapy.

Claims

1. A method for obtaining monocytes or NK cells, comprising:

(a) collecting a monocyte sample by separating the monocytes from peripheral blood by using an antibody specific for a surface marker of the monocytes, wherein the peripheral blood is obtained from a human;

(b) collecting, as an NK cell sample, cell populations remaining after the separation of the monocytes; and

(c) growing the monocytes or the NK cells by culturing at least one of the monocyte sample and the NK cell sample,

wherein in the step (c), at least the NK cells are grown by culturing the NK cell sample without carrying out further separation steps by use of a second medium containing an anti-CD56 antibody.

2. The method as set forth in claim 1, wherein the second medium further contains IL-2, IL-12, and IL-15.

3. The method as set forth in claim 1, wherein in the step (c), the monocytes are further grown by culturing the monocyte sample by use of a first medium containing Flt-3L.