# US 8,569,060 B2 · Method of producing a population of cells > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,569,060 B2 - Document ID: US8569060B2 - Status: Granted - Kind: B2 - Application number: 12/812,143 - Filed: 2009-01-08 - Published: 2010-11-11 - Granted: 2013-10-29 - Earliest claimed: 2008-01-08 - Assignees: The University of Queensland - Applicants: Nicholas Eion Timmins; Lars Keld Nielsen; Emma Louise Palfreyman - Inventors: Nicholas Eion Timmins; Lars Keld Nielsen; Emma Louise Palfreyman - Web page: https://patentgrep.com/patent/US8569060B2 ## CPC - A61K 35/18: Erythrocytes (haemoglobin A61K38/42) - A61K 35/15: 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) - A61K 35/17: Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes (when activated by a specific antigen A61K39/00) - A61K 35/19: Platelets; Megacaryocytes - C12N 5/0641: {Erythrocytes} - C12N 2500/24: Iron; Fe chelators; Transferrin - C12N 2500/25: Insulin-transferrin; Insulin-transferrin-selenium - C12N 2501/125: Stem cell factor [SCF], c-kit ligand [KL] - C12N 2501/14: Erythropoietin [EPO] - C12N 2501/23: Interleukins [IL] - C12N 2501/39: Steroid hormones - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract An in vitro or ex vivo method of producing a population of lineage committed haematopoietic progenitor or mature haematopoietic cells other than cells of the neutrophil lineage, including the steps of providing a population of haematopoietic progenitor cells; and culturing the haematopoietic progenitor cells in an animal cell culture medium including one or more cytokines that differentiate the haematopoietic progenitor cells into lineage committed haematopoietic progenitor and/or mature haematopoietic cells, under static conditions until the cells are at a cell density at which oxygen transfer via the surface of the culture medium is insufficient for growth of the progenitor cells and progeny thereof under static conditions, and then agitating the culture medium to produce a population of lineage committed haematopoietic progenitor or mature haematopoietic cells other than cells of the neutrophil lineage. ## Claims ### Claim 1 An in vitro or ex vivo method of producing a population of lineage committed haematopoietic progenitor or mature haematopoietic cells other than cells of the neutrophil lineage, which method comprises the steps of: (a) providing a population of cells comprising haematopoietic progenitor cells; and (b) culturing the population of cells in an animal cell culture medium comprising one or more cytokines that differentiate said progenitor cells into lineage committed haematopoietic progenitor and/or mature haematopoietic cells, under static conditions until the cells are at a cell density at which oxygen transfer via the surface of the culture medium is insufficient for growth of the progenitor cells and progeny thereof under static conditions, and then agitating the culture medium thereafter, to produce a population of lineage committed haematopoietic progenitor or mature haematopoietic cells other than cells of the neutrophil lineage. ### Claim 2 (depends on 1) The method of claim 1, wherein the culture medium is agitated once the total cell density is at least about 100,000 to about 200,000 cells per ml. ### Claim 3 (depends on 1) The method of claim 1, wherein the population of cells of step (b) is at an initial cell density of less than about 20,000 haemotopoietic progenitor cells per ml. ### Claim 4 (depends on 1) The method of claim 1, wherein the initial culture medium of step (b) further comprises cells other than haematopoietic progenitor cells such that the total initial cell density is at least about 100,000 cells per ml of medium. ### Claim 5 (depends on 4) The method of claim 4, wherein the cells other than haematopoietic progenitor cells are peripheral blood mononuclear cells. ### Claim 6 (depends on 1) The method of claim 1, wherein the cells are cultured in a collapsible culture vessel and wherein the culture vessel is partly or fully inflated and the agitation of the vessel generates a wave motion in the culture medium. ### Claim 7 (depends on 1) The method of claim 1, wherein the population of haematopoietic progenitor cells has been enriched. ### Claim 8 A method of increasing the number of haematopoietic cells in a patient, the method comprising: (a) providing a population of cells comprising haematopoietic progenitor cells; (b) culturing the population of cells in an animal cell culture medium comprising one or more cytokines that differentiate said progenitor cells into lineage committed haematopoietic progenitor and/or mature haematopoietic cells, under static conditions until the cells are at a cell density at which oxygen transfer via the surface of the culture medium is insufficient for growth of the progenitor cells and progeny thereof under static conditions, and then agitating the culture medium thereafter, to produce a population of lineage committed haematopoietic progenitor or mature haematopoietic cells other than cells of the neutrophil lineage; and (c) administering to the patient the population of lineage committed haematopoietic progenitor or mature haematopoietic cells other than cells of the neutrophil lineage obtained in step (b) such that the number of haematopoietic cells in the patient is increased. ### Claim 9 (depends on 8) The method of claim 8, wherein the population of cells comprising haematopoietic progenitor cells is obtained from the patient. ### Claim 10 (depends on 1) The method of claim 1, wherein the population of haematopoietic progenitor cells is provided as a non-enriched population of mononuclear cells.