Mesenchymal precursor cell

• US 8,158,118 B2
• Filed: 06/29/2005
• Issued: 04/17/2012
• Est. Priority Date: 07/07/1999
• Status: Expired due to Term

• Current Assignee: Mesoblast, Inc. (Mesoblast Ltd.)
• Original Assignee: Angioblast Systems, Inc.
• Inventors: Simmons, Paul, Zannettino, Andrew, Gronthos, Stan
• Primary Examiner(s): BELYAVSKYI, MICHAIL A
• Application Number:
  US11/169,875
• Publication Number:
  US 20050281790A1
• Time in Patent Office: 2,484 Days
• Field of Search: None
• US Class Current: 424/93.1
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: C12N 2500/42 Organic phosphate, e.g. beta glycerophosphate
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 5/0663 Bone marrow mesenchymal stem cells (BM-MSC)

Abstract

A method of enriching mesenchymal precursor sells including the step of enriching for cells based on at least two markers. The markers may be either i) the presence of markers specific for mesenchymal precursor cells, ii) the absence of markers specific for differentiated mesenchymal cells, or iii) expression levels of markers specific for mesenchymal precursor cells. The method may include a first solid phase sorting step utilizing MACS recognizing expression of the antigen to the STRO-1 Mab, followed by a second sorting step utilizing two color FACS to screen for the presence of high level STRO-1 antigen expression as well as the expression of VCAM-1.

Claims

1. A purified population of STRO-1 positive cells wherein greater than 5% of the STRO-1 positive cells in the population are capable of giving rise to colony forming unit-fibroblasts (CFU-F).

2. The population of STRO-1 positive cells of claim 1, wherein greater than 10% of the STRO-1 positive cells are capable of giving rise to colony forming unit-fibroblasts (CFU-F).

3. The population of STRO-1 positive cells of claim 1, wherein greater than 40% of the STRO-1 positive cells are capable of giving rise to colony forming unit-fibroblasts (CFU-F).

4. The population of STRO-1 positive cells of claim 1, wherein greater than 50% of the STRO-1 positive cells are capable of giving rise to colony forming unit-fibroblasts (CFU-F).

5. The population of STRO-1 positive cells of claim 1, wherein the cells carry at least one additional marker selected from the group of surface markers consisting of THY-1, VCAM-1, STRO-2, and CD146.

6. The population of STRO-1 positive cells of claim 5, wherein the cells carry the marker VCAM-1.

7. The population of STRO-1 positive cells of claim 1, wherein said STRO-1 positive cells are capable of differentiation into at least two committed cell types selected from the group consisting of adipose, areolar, osseous, cartilaginous, elastic, and fibrous connective tissue.

8. The population of STRO-1 positive cells of claim 1, wherein the population is suitable for seeding onto a vehicle for implantation to assist in bone growth.

9. The population of STRO-1 positive cells of claim 1, wherein said STRO-1 positive cells comprise an exogenous nucleic acid that expresses a therapeutic agent transformed into them such that the population of cells may be introduced into the body of a patient to release the therapeutic agent.

10. The population of STRO-1 positive cells of claim 1, wherein the population is used to augment bone marrow transplantation.

11. The population of STRO-1 positive cells of claim 1, wherein the STRO-1 positive cells are negative for at least one marker selected from the group consisting of CBFA-1, collagen type II, PPARγ

2, and glycophorin A.

12. A composition comprising a purified population of STRO-1 positive cells wherein greater than 5% of the STRO-1 positive cells are capable of giving rise to colony forming unit-fibroblasts (CFU-F) and haemopoietic cells.

13. The composition of claim 12, wherein the population of STRO-1 positive cells is preadsorbed onto a ceramic vehicle that is precoated with fibronectin and is suitable for implantation to augment bone marrow transplantation.

14. The composition of claim 12, wherein said STRO-1 positive cells comprise an exogenous nucleic acid that expresses a therapeutic agent transformed into them such that the composition may be introduced into the body of a patient to release the therapeutic agent.