Differentiated pluripotent stem cell progeny depleted of extraneous phenotypes

• US 9,074,182 B2
• Filed: 11/16/2012
• Issued: 07/07/2015
• Est. Priority Date: 06/25/2009
• Status: Active Grant

• Current Assignee: Asterias Biotherapeutics Incorporated (Lineage Cell Therapeutics, Inc.)
• Original Assignee: Asterias Biotherapeutics Incorporated (Lineage Cell Therapeutics, Inc.)
• Inventors: Lebkowski, Jane S., Priest, Catherine Ann, Okamura, Ross M.
• Primary Examiner(s): Schultz, Doug
• Application Number:
  US13/679,663
• Publication Number:
  US 20130210109A1
• Time in Patent Office: 963 Days
• US Class Current: 1/1
• CPC Class Codes: C12N 2502/09 epidermal cells, skin cells, oral mucosa cells
• CPC Class Codes: C12N 2502/23 Gastro-intestinal tract cells
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 5/0081 Purging biological preparations of unwanted cells
• CPC Class Codes: C12N 5/0607 Non-embryonic pluripotent stem cells, e.g. MASC induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0622 Glial cells, e.g. astrocytes, oligodendrocytes; Schwann cells
• CPC Class Codes: C12N 5/0623 Stem cells

Abstract

The invention provides methods for depleting extraneous phenotypes from a mixed population of cells comprising the in vitro differentiated progeny of primate pluripotent stem cells. The invention also provides mixed cell populations enriched for a target cell phenotype where the mixed cell population comprises the differentiated in vitro progeny of primate embryonic stem cells.

Claims

1. A method of reducing the number of cells co-expressing TRA-1-60 and EpCAM in a mixed population of cells comprising:

a) providing a mixed population of cells comprising the in vitro progeny of pPS cells, wherein the mixed population of cells comprises i) cells co-expressing TRA-1-60 and EpCAM, and ii) oligodendrocytes differentiated from the pPS cells;

b) contacting the mixed population of cells with one or more ligands that specifically bind to EpCAM expressed on cells of the mixed population of cells; and

c) separating the ligand bound EpCAM expressing cells from the rest of the mixed population of cells to thereby reduce the number of cells expressing TRA-1-60 in the mixed population of cells.

2. The method of claim 1, wherein the ligand is an antibody.

3. The method of claim 1, wherein the ligand is bound to a solid support.

4. The method of claim 3, wherein the solid support is a bead.

5. The method of claim 4, wherein the bead is a magnetic bead.

6. The method of claim 1, wherein separating the ligand bound EpCAM expressing cells from the rest of the mixed population of cells is performed by applying an external force to the mixed population of cells.

7. The method of claim 6, wherein the external force is provided by a magnetic field.