PROGENITOR CELL POPULATIONS, EXPANSION THEREOF, AND GROWTH OF NON-HEMATOPOIETIC CELL TYPES AND TISSUES THEREFROM

• US 20080020459A1
• Filed: 07/26/2007
• Published: 01/24/2008
• Est. Priority Date: 04/24/2001
• Status: Active Grant

• Current Assignee: Dolores Baksh, John Davies, Peter Zandstra
• Original Assignee: Dolores Baksh, John Davies, Peter Zandstra
• Inventors: DAVIES, John, ZANDSTRA, Peter, BAKSH, Dolores
• Granted Patent: US 9,644,182 B2
• Time in Patent Office: Days
• US Class Current: 435/372
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C12N 2501/125 Stem cell factor [SCF], c-kit ligand [KL]
• CPC Class Codes: C12N 2501/23 Interleukins [IL]
• CPC Class Codes: C12N 5/0663 Bone marrow mesenchymal stem cells (BM-MSC)

Abstract

Described is a method of expanding human progenitor cells by suspension culturing under non-static conditions. The culturing method provides a three-dimensional space for the rapid expansion of desirable progenitors. By this method, a new compartment of multipotential progenitor cells has been identified, which give rise under differentiation conditions to progeny including osteoblasts, chondrocytes, myoblasts, adipocytes, and other non-hematopoietic cell types. Their use in cell and tissue-based engineering is described.

Claims

1. An enriched progenitor cell population comprising non-hematopoietic progenitor cells extractable from bone marrow, wherein the cell population is substantially devoid of at least one type of hematopoietic progenitor cell.

2. An enriched progenitor cell population according to claim 1, characterized by the absence of at least one hematopoietic progenitor cell type, wherein said cell type is one having a surface marker selected from CD3, CD 14, CD38, CD45, CD66, CD119.

3. An enriched progenitor cell population according to claim 1, wherein the population is substantially devoid of at least one of the progenitor cell types bearing an MHC determinant selected from HLA-A, HLA-B, HLA-C, HLA-DR, HLA-DP and HLA-DQ.

4. An enriched progenitor cell population according to claim 2, wherein the population is substantially devoid of progenitor cell types that bear any MHC determinant selected from HLA-A, HLA-B, HLA-C, HLA-DR, HLA-DP and HLA-DQ.

7. The method according to claim 5, wherein said progenitor cell population is an enriched progenitor cell population according to any one of claims 1-4.

15. An expanded progenitor cell population, prepared by the suspension culturing of an enriched progenitor cell population according to any one of claims 1-4.

16. A composition, useful for delivering non-hematopoietic progenitor cells to an environment conducive to the formation of differentiated cells therefrom, the composition comprising an expanded population of human progenitor cells according to any one of claims 1-4, 14 or 15, and a vehicle for delivering said cells to said environment.

17. A progenitor cell population as claimed in any one of claims 1-4, 14 and 15, in combination with one or more factors for inducing the differentiation thereof.

5. A method for expanding human non-hematopoietic progenitor cells, the method comprising the step of subjecting a progenitor cell population comprising non-hematopoietic progenitor cells to non-static suspension culturing in a suitable medium for a period of time effective to expand said progenitor cells.

6. The method according to claim 5 wherein said progenitor cell population comprises human mesenchymal progenitor cells.

8. The method according to any one of claims 5-7, wherein said growth medium includes growth factors for human mesenchymal progenitor cells.

9. The method according to any one of claims 5-7, wherein the growth medium excludes at least one factor specific for the growth of hematopoietic progenitor cells.

10. The method according to claim 8 or claim 9 wherein said growth factors are cytokines.

11. The method according to claim 10 wherein said cytokines are a mixture of stem cell factor and interleukin-3.

12. The method according to any one of claims 5-10, wherein said suspension is stirred during said effective period of time.

13. The method according to claim 11, wherein said suspension is maintained at a temperature of about 37°

C. during said effective period of time.

14. A population of expanded human progenitor cells, characterized in that:

1) when cultured under conditions appropriate for differentiation, said population is able gives rise to differentiated cell types that include bone cells, fibroblast cells and neuronal cells;

2) the population has been expanded in suspension culture;

3) the progenitor cells within the population have not been selected on the basis of adherence;

4) the population phenotype is SH4 negative; and

5) the population results from culturing in the absence of at least one culture supplement specific for hematopoietic progenitor cell maintenance.

18. A method for producing non-hematopoietic cells, comprising the steps of subjecting a progenitor cell population comprising non-hematopoietic progenitor cells to non-static suspension culturing in a suitable medium for a period of time effective to expand said progenitor cells, and then delivering the expanded progenitor cells or a subpopulation thereof to an environment conducive to the differentiation thereof into the desired cell type.

22. The method according to claim 18, wherein the expanded progenitor cells or a subpopulation thereof are delivered to an environment conducive to the differentiation thereof into myoblasts.

23. The method according to claim 18, wherein the expanded progenitor cells or a subpopulation thereof are delivered to an environment conducive to the differentiation thereof into neural cells.

24. The method according to claim 18, wherein the expanded progenitor cells or a subpopulation thereof are delivered to an environment conducive to the differentiation thereof into adipocytes.

25. The method according to claim 18, wherein the expanded progenitor cells or a subpopulation thereof are delivered to an environment conducive to the differentiation thereof into endothelial cells.

19. A method of providing cells differentiated from mesenchymal progenitor cells, comprising the steps of subjecting a progenitor cell population comprising mesenchymal progenitor cells to non-static suspension culturing in a suitable medium for a period of time effective to expand said progenitor cells, and then delivering the expanded progenitor cells or a subpopulation thereof to an environment conducive to the differentiation thereof into a mesenchymal cell type.

20. The method according to claim 19, wherein the expanded progenitor cells or a subpopulation thereof are delivered to an environment conducive to the differentiation thereof into bone cells.

21. The method according to claim 19, wherein the expanded progenitor cells or a subpopulation thereof are delivered to an environment conducive to the differentiation thereof into chondrocytes.

26. A cell population representing a new compartment of progenitor cells, said cell population comprising progenitor cells which, under differentiation conditions give rise to a variety of non-hematopoietic cell types including osteoblasts, adipocytes and myoblasts, wherein said cell population results from the non-static suspension culturing of an input population of progenitor cells obtained without prior selection based on cell adherence.