Mesenchymal precursor cell

• US 7,122,178 B1
• Filed: 07/07/2000
• Issued: 10/17/2006
• Est. Priority Date: 07/07/1999
• Status: Expired due to Term

• Current Assignee: Mesoblast, Inc. (Mesoblast Ltd.)
• Original Assignee: Angioblast Systems, Inc.
• Inventors: Gronthos, Stan, Zannettino, Andrew, Simmons, Paul
• Primary Examiner(s): Chan, Christina
• Assistant Examiner(s): Belyavskyi, Michail A.
• Application Number:
  US10/030,411
• Time in Patent Office: 2,293 Days
• Field of Search: 435/325, 435/372, 424/93.1, 424/577
• 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 population of cells enriched for STRO-1^bright cells, wherein such STRO-1^bright cells are mesenchymal precursor cells which comprise mesenchymal precursor cells capable of giving rise to colony forming unit-fibroblasts (CFU-F).

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

3. The enriched population of cells as in claim 2, wherein the mesenchymal precursor cells carry the markers STRO-1 and VCAM-1.

4. The enriched population of cells as in claim 1, wherein said STRO-1^bright 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.

5. The enriched population of cells as in claim 1, wherein the enriched population is suitable for seeding onto a vehicle for implantation to assist in bone growth.

6. The enriched population of cells as in claim 1, wherein said STRO-1^bright cells in the enriched population 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.

7. The enriched population of cells as in claim 1, wherein the enriched population is used to augment bone marrow transplantation.

8. A composition comprising the enriched population of cells of claim 1.

9. The composition as in claim 8, wherein the composition is preadsorbed onto a ceramic vehicle that is precoated with fibronectin and is suitable for implantation to augment bone marrow transplantation.

10. The composition as in claim 8, wherein the composition is suitable for use in augmenting bone marrow transplantation.

11. The composition as in claim 8, wherein the composition also comprises haemopoietic cells.

12. The composition as in claim 8, wherein said STRO-1^bright cells in the enriched population 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.

13. The enriched population of cells as in claim 1, wherein the STRO-1^bright cells are negative for at least one marker selected from the group consisting of CBFA-1, collagen type II, PPARγ

2, and glycophorin A.