Differentiated pluripotent stem cell progeny depleted of extraneous phenotypes

• US 8,323,966 B2
• Filed: 06/25/2010
• Issued: 12/04/2012
• Est. Priority Date: 06/25/2009
• Status: Active Grant

• Current Assignee: Asterias Biotherapeutics Incorporated (Lineage Cell Therapeutics, Inc.)
• Original Assignee: Geron Corporation
• Inventors: Lebkowski, Jane S., Priest, Catherine A., Okamura, Ross M.
• Primary Examiner(s): Schultz, Doug
• Application Number:
  US12/823,739
• Publication Number:
  US 20100330670A1
• Time in Patent Office: 893 Days
• Field of Search: None
• US Class Current: 435/368
• 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 extraneous phenotypic cells in a mixed population of cells comprising a) contacting the mixed population of cells with one or more ligands that specifically bind to the extraneous phenotypic cells;

and b) separating the ligand bound extraneous phenotypic cells from the rest of the mixed population of cells thereby reducing the number of extraneous phenotypic cells from a mixed population of cells, wherein the mixed population of cells comprises the in vitro differentiated progeny of pPS cells including the extraneous phenotypic cells and wherein the ligand is an antibody that binds specifically to EpCAM and the mixed population of cells comprises a targeted phenotype and the targeted phenotype is an oligodendrocyte.

2. The method of claim 1, wherein the extraneous phenotypic cell is an epithelial cell.

3. The method of claim 2, wherein the epithelial cell expresses a cytokeratin.

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

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

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

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

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