Differentiated cells suitable for human therapy

• US 20060134782A1
• Filed: 02/21/2006
• Published: 06/22/2006
• Est. Priority Date: 11/27/2000
• Status: Active Grant

• Current Assignee: Asterias Biotherapeutics Incorporated (Lineage Cell Therapeutics, Inc.)
• Original Assignee: Asterias Biotherapeutics Incorporated (Lineage Cell Therapeutics, Inc.)
• Inventors: Gold, Joseph D., Lebkowski, Jane S.
• Granted Patent: US 8,426,198 B2
• Time in Patent Office: Days
• US Class Current: 435/366
• 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 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 2502/13 connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts"
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0606 Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696

Abstract

This invention provides a system for producing differentiated cells from a stem cell population for use wherever a relatively homogenous cell population is desirable. The cells contain an effector gene under control of a transcriptional control element (such as the TERT promoter) that causes the gene to be expressed in relatively undifferentiated cells in the population. Expression of the effector gene results in depletion of undifferentiated cells, or expression of a marker that can be used to remove them later. Suitable effector sequences encode a toxin, a protein that induces apoptosis; a cell-surface antigen, or an enzyme (such as thymidine kinase) that converts a prodrug into a substance that is lethal to the cell. The differentiated cell populations produced according to this disclosure are suitable for use in tissue regeneration, and non-therapeutic applications such as drug screening.

Claims

1. A pair of isolated cell populations, consisting of:

a first cell population comprising undifferentiated cells from a line of human embryonic stem (hES) cells; and

a second cell population that is free of undifferentiated hES cells, but which contains progeny of said hES cell line.

2. The cell populations of claim 1, wherein the cells contain a nucleic acid molecule comprising the structure P-X, wherein:

X is a nucleic acid sequence encoding a product that is lethal to a cell in which it is expressed, or renders a cell in which it is expressed susceptible to a lethal effect of an external agent; and

P is a transcriptional control element that causes X to be preferentially expressed in undifferentiated cells.

3. The cell populations of claim 2, wherein X encodes a toxin, or a protein that induces or mediates apoptosis.

4. The cell populations of claim 2, wherein X encodes an enzyme that converts a prodrug to a compound that is lethal to a cell in which X is expressed.

5. The cell populations of claim 4, wherein X encodes a thymidine kinase.

6. The cell populations of claim 2, wherein P-X is an introduced heterologous molecule.

7. The cell populations of claim 2, wherein P is an endogenous transcriptional control element.

8. The cell populations of claim 2, wherein P is an OCT-4 promoter or a promoter of telomerase reverse transcriptase (TERT).

9. The cell populations of claim 1, wherein the second cell population is a population of neurons or neural precursor cells.

10. The cell populations of claim 1, wherein the second cell population is a population of hepatocytes.

11. The cell populations of claim 1, wherein the second cell population is a population of cardiomyocytes.

12. The cell populations of claim 1, wherein the second cell population is formulated for tissue reconstitution or regeneration in a human patient.