Perfusion bioreactor for culturing CD200—placenta adherent cells

• US 9,758,762 B2
• Filed: 08/20/2013
• Issued: 09/12/2017
• Est. Priority Date: 09/02/2008
• Status: Active Grant

• Current Assignee: Pluri Biotech Limited
• Original Assignee: Pluristem Ltd. (Pluri Inc.)
• Inventors: Meiron, Moran, Toren, Amir, Ofir, Racheli, Drori-Carmi, Nirit
• Primary Examiner(s): Moloye, Titilayo
• Application Number:
  US13/971,013
• Publication Number:
  US 20130337558A1
• Time in Patent Office: 1,484 Days
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61P 19/04 for non-specific disorders of the connective tissue
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/04 specific for metastasis
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 2500/34 Sugars
• CPC Class Codes: C12N 5/0605 Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
• CPC Class Codes: C12N 5/0653 Adipocytes; Adipose tissue

Abstract

A method of culturing adherent cells from a placenta or adipose tissue is disclosed. The method comprises culturing the adherent cells from the placenta or adipose tissue under 3 dimensional (3D) culturing conditions which allow cell expansion, the conditions comprising perfusion.

Claims

1. A bioreactor comprising:

a perfusion chamber housing;

i) a population of adherent cells from placenta that are negative for the marker CD200, wherein the adherent cells from placenta do not exhibit osteogenic differentiation when subjected to conditions that induce bone marrow cells to undergo osteogenic differentiation, wherein the adherent cells from placenta are present in the perfusion chamber under culturing conditions which allow for cell expansion, said conditions comprising perfusion of a cell culture medium through the bioreactor, and wherein the bioreactor is configured to adjust the rate of said perfusion according to the glucose concentration of the culture medium, and

ii) a plurality of 3D porrosive carriers on which the cells are cultured within the bioreactor, wherein each porrosive carrier comprises a fiber matrix, and wherein the carriers are packed in the perfusion chamber.

2. The bioreactor of claim 1, wherein more than 25% of the adherent cells from placenta are in S and G2/M cell cycle phases.

3. The bioreactor of claim 1, wherein the adherent cells from placenta are less committed to differentiation into adipogenic lineages as compared to adherent cells from bone marrow grown under the same conditions.

4. The bioreactor of claim 1, wherein the fiber matrix is composed of a non-woven fabric matrix.

5. The bioreactor of claim 4, wherein the non-woven fabric matrix comprises a polyester.

6. A bioreactor comprising:

a perfusion chamber configured for housing a population of adherent cells from placenta that are negative for the marker CD200 under culturing conditions which allow for cell expansion, said conditions comprising perfusion of a cell culture medium through the bioreactor, wherein the bioreactor is configured to adjust the rate of said perfusion according to the glucose concentration of the culture medium, and wherein the adherent cells from placenta do not exhibit osteogenic differentiation when subjected to conditions that induce bone marrow cells to undergo osteogenic differentiation, wherein the adherent cells from placenta are present in the perfusion chamber.