Isolation of adult multipotential cells by tissue non-specific alkaline phosphatase
View Patent ↗The present invention relates to the use of tissue non-specific alkaline phosphatase (TNAP) as a marker for identifying and/or isolating adult multipotential cells. The present invention also relates to cell populations enriched by methods of the present invention and therapeutic uses of these cells.
1. A method of enriching for adult STRO-1 + multipotential cells, the method comprising:
(i) preparing a cell sample from a tissue source comprising cells that express the marker tissue non-specific alkaline phosphatase (TNAP) and enriching for cells that express the TNAP marker and CD45; and
(ii) enriching the cell population obtained in step (i) for adult multipotential cells expressing the STRO-1 marker.
2. A method according to claim 1 which comprises:
contacting the cell sample with a TNAP binding agent under conditions that allows binding of TNAP to the TNAP binding agent; and separating cells bound to the TNAP binding agent.
3. A method according to claim 2 wherein the TNAP binding agent binds specifically to the BAP isoform of TNAP.
4. A method according to claim 2 wherein the TNAP binding agent is an antibody that binds to the same epitope as the STRO-3 antibody produced by the hybridoma cell line deposited with ATCC on 19 Dec. 2005 under the provisions of the Budapest Treaty under deposit accession number PTA-7282.
5. A method according to claim 4 wherein the antibody is the STRO-3 antibody produced by the hybridoma cell line deposited with ATCC on 19 Dec. 2005 under the provisions of the Budapest Treaty under deposit accession number PTA-7282.
6. A method according to claim 1 for identifying the presence of an adult multipotential cell in a cell sample, the method comprising identifying cells in the sample that express the TNAP marker.
7. An enriched population of adult multipotential cells obtained by a method according to claim 1 .
8. An expanded cell population obtained by culturing an enriched population of adult multipotential cells according to claim 7 .
9. A composition comprising an expanded cell population of claim 8 .
10. A method of generating a tissue specific committed cell population, the method comprising culturing a population of adult multipotential cells according to claim 7 in the presence of one or more stimulatory factors; and subjecting said cultured population to conditions biasing differentiation of the adult multipotential cells to a specific tissue type.
11. A composition comprising a population of enriched adult multipotential cells according to claim 7 .
12. An enriched population of adult multipotential cells obtained by a method according to claim 1 , wherein at least 1% of the total enriched cell population are STRO-1 + ′ TNAP + adult multipotential cells.
13. An expanded cell population obtained by culturing an enriched population of adult multipotential cells according to claim 12 .
14. A method of generating a tissue specific committed cell population, the method comprising culturing a population of adult multipotential cells according to claim 12 in the presence of one or more stimulatory factors; and subjecting said cultured population to conditions biasing differentiation of the adult multipotential cells to a specific tissue type.
15. A composition comprising a population of enriched adult multipotential cells according to claim 12 .
16. An isolated cell which has been obtained by a method according to claim 1 , or a progeny cell thereof, wherein the cell is genetically modified.
17. A method according to claim 1 , wherein step (ii) results in loss of expression of CD45.
18. A method according to claim 1 , wherein step (ii) comprises culturing the cell population obtained from step (i).