Mesenchymal precursor cell
View Patent ↗A method of enriching mesenchymal precursor cells 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 utilising MACS recognising expression of the antigen to the STRO-1 Mab, followed by a second sorting step utilising two colour FACS to screen for the presence of high level STRO-1 antigen expression as well as the expression of VCAM-1.
1. A process for making a population of mesenchymal precursor cells comprising
obtaining a population of cells comprising cells which express STRO-1 at a higher level and cells which express STRO-1 at a lower level, relative to each other, and
enriching the population for cells which express STRO-1 at the higher level relative to cells which express STRO-1 at the lower level, thereby making the population of mesenchymal precursor cells.
2. The process of claim 1 , wherein the cells which express STRO-1 at the higher level also express at least one other antigen selected from the group consisting of LFA-3, ICAM-1, PECAM-1, P-selectin, L-selectin, CD49b/CD29, CD49c/CD29, CD49d/CD29, CD29, CD18, CD61, 6-19, thrombomodulin, telomerase, CD10, CD13, integrin beta, and SCF.
3. The process of claim 1 , wherein the enriching step is performed so as to make a cell population in which at least 1% of the cells are capable of giving rise to colony forming units-fibroblast (CFU-F).
4. The process of claim 1 , wherein the enriching step is performed so as to make a cell population in which at least 5% of the cells are capable of giving rise to colony forming units-fibroblast (CFU-F).
5. The process of claim 1 , wherein the enriching step is performed so as to make a cell population in which at least 10% of the cells are capable of giving rise to colony forming units-fibroblast (CFU-F).
6. The process of claim 1 , wherein the enriching step is performed so as to make a cell population in which at least 40% of the cells are capable of giving rise to colony forming units-fibroblast (CFU-F).
7. The process of claim 1 , wherein the enriching step is performed so as to make a cell population in which at least 2% of the cells carry both the antigen identified by STRO-1 and VCAM-1.
8. The process of claim 1 , wherein the enriching step is performed so as to make a cell population in which cells which express STRO-1 at the higher level are negative for at least one marker selected from the group consisting of CBFA-1, collagen type II, PPARγ2, osteopontin, osteocalcin, parathyroid hormone receptor, leptin, human adipocyte lipid binding protein (H-ALBP), aggrecam, Ki67, and glycophorin A.
9. The process of claim 1 , wherein the enriching step is performed so as to make a cell population in which at least 1.5% of the cells in the population express STRO-1 at the higher level.
10. The process of claim 1 , wherein the enriching step is performed so as to make a cell population in which at least 2% of the cells in the population express STRO-1 at the higher level.
11. The process of claim 1 , wherein the population of cells is stromal stem cells obtained from bone marrow, blood, epidermis, or hair follicles.
12. The process of claim 11 , wherein the population of cells is obtained from bone marrow.
13. The process of claim 1 , wherein obtaining the population of cells comprises harvesting the population of the cells from a subject.
14. The process of claim 1 , wherein the cells which express STRO-1 at the higher level also express at least one other marker selected from VACM-1, THY-1, CD146, and STRO-2.
15. The process of claim 1 , wherein the enriching step comprises a separation step based on recognition of cells expressing the STRO-1 marker.
16. The process of claim 15 , wherein the recognition of cells expressing the STRO-1 marker is achieved by a binding agent which binds to a marker on the surface of the cells expressing the STRO-1 marker.
17. The process of claim 16 , wherein the binding agent is a monoclonal antibody or molecule based on a monoclonal antibody.
18. The process of claim 1 , wherein at least a proportion of the population of mesenchymal precursor cells is capable of differentiation into at least two committed cell types each selected from the group consisting of adipose, areolar, osseous, cartilaginous, elastic, and fibrous connective tissue.
19. The process of claim 1 , further comprising transforming the cell population with an exogenous nucleic acid.