Neutral progenitor cells from hippocampal tissue and a method for isolating and purifying them

• US 20030211087A1
• Filed: 10/23/2002
• Published: 11/13/2003
• Est. Priority Date: 01/18/2000
• Status: Active Grant

• Current Assignee: Cornell Research Foundation Incorporated (Cornell University)
• Original Assignee: Cornell Research Foundation Incorporated (Cornell University)
• Inventors: Goldman, Steven A.
• Granted Patent: US 7,785,882 B2
• Time in Patent Office: Days
• US Class Current: 424/93.21
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 2510/04 Immortalised cells
• CPC Class Codes: C12N 2830/002 inducible enhancer/promoter combination, e.g. hypoxia, iron, transcription factor
• CPC Class Codes: C12N 2830/008 cell type or tissue specific enhancer/promoter combination
• CPC Class Codes: C12N 2830/30 being an enhancer not forming part of the promoter region
• CPC Class Codes: C12N 2830/85 mammalian
• CPC Class Codes: C12N 5/0623 Stem cells

Abstract

The present invention relates to an enriched or purified preparation of isolated hippocampal neural progenitor cells and progeny thereof. The present invention also relates to a method of separating neural progenitor cells from a mixed population of cell types from hippocampal tissue. This method includes selecting a promoter which functions selectively in the neural progenitor cells, introducing a nucleic acid molecule encoding a fluorescent protein under control of said promoter into all cell types of the mixed population of cell types from hippocampal tissue, allowing only the neural progenitor cells, but not other cell types, within the mixed population to express said fluorescent protein, identifying cells of the mixed population of cell types that are fluorescent, which are restricted to the neural progenitor cells, and separating the fluorescent cells from the mixed population of cell types, wherein the separated cells are restricted to the neural progenitor cells.

Claims

1. An enriched or purified preparation of isolated hippocampal neural progenitor cells and progeny thereof.

2. An enriched or purified preparation of isolated hippocampal neural progenitor cells and progeny thereof according to claim 1 which are human.

3. An enriched or purified preparation of isolated hippocampal neural progenitor cells and progeny thereof according to claim 2 which are from an adult.

4. An enriched or purified preparation of isolated hippocampal neural progenitor cells and progeny thereof according to claim 2 which are from a fetus.

5. An enriched or purified preparation of isolated hippocampal neural progenitor cells and progeny thereof according claim 1 which gives rise to dentate gyrus.

6. An enriched or purified preparation of isolated hippocampal neural progenitor cells and progeny thereof according claim 1, wherein the neural progenitor cells or progeny thereof are immortalized.

7. An enriched or purified preparation of isolated hippocampal neural progenitor cells and progeny thereof according claim 6, wherein the neural progenitor cells or progeny thereof are immortalized with a transforming oncogene.

8. An enriched or purified preparation of isolated hippocampal neural progenitor cells and progeny thereof according claim 1, wherein the neural progenitor cells or progeny thereof are transduced with an exogenous transgene.

9. A method of separating neural progenitor cells from a mixed population of cell types from hippocampal tissue, said method comprising:

selecting a promoter which functions selectively in the neural progenitor cells;

introducing a nucleic acid molecule encoding a fluorescent protein under control of said promoter into all cell types of the mixed population of cell types from hippocampal tissue;

allowing only the neural progenitor cells, but not other cell types, within the mixed population to express said fluorescent protein;

identifying cells of the mixed population of cell types that are fluorescent, which are restricted to the neural progenitor cells; and

separating the fluorescent cells from the mixed population of cell types, wherein the separated cells are restricted to the neural progenitor cells.

10. A method according to claim 9, wherein the mixed population of cell types are mammalian.

11. A method according to claim 10, wherein the mixed population of cell types are human.

12. A method according to claim 11, wherein the mixed population of cell types are from an adult.

13. A method according to claim 11, wherein the mixed population of cell types are from a fetus.

14. A method according to claim 9, wherein the promoter is a Tα

1 tubulin promoter.

15. A method according to claim 9, wherein the promoter is a nestin enhancer.

16. A method according to claim 9, wherein the promoter is a Sox2 promoter.

17. A method according to claim 9, wherein said introducing comprises viral mediated transformation of the mixed population of cell types.

18. A method according to claim 17, wherein said viral mediated transformation comprises adenovirus mediated transformation.

19. A method according to claim 9, wherein said introducing comprises electroporation.

20. A method according to claim 9, wherein said introducing comprises liposomal mediated transformation of said plurality of cells.

21. A method according to claim 9, wherein said separating comprises fluorescence activated cell sorting.

22. A method according to claim 9, wherein the mixed population of cell types is in cell culture.

23. A method according to claim 9 further comprising:

transplanting the separated cells into a subject.