Stem cell aggregates and methods for making and using
View Patent ↗The invention is directed to compositions of cell aggregates and methods for making and using the cell aggregates where the aggregates comprise cells that are not embryonic stem cells but can differentiate into cell types of at least two of ectodermal, undo dermal, and mesodermal embryonic germ layers, e.g., stem cells.
1. A method comprising (1) disaggregating cells in an aggregate, the aggregate having been formed by exposing cells that are not embryonic stem cells, embryonic germ cells, or germ cells and can differentiate into cell types of at least two of the endodermal, ectodermal, mesodermal embryonic lineages, to conditions under which the cells aggregate, wherein the conditions comprise an oxygen concentration of about 1-10%, wherein the aggregate from which the disaggregated cells are derived does not originate from two-dimensional culture and (2) administering to a subject, in an amount and for a time sufficient to achieve a desired effect in the subject, the cells derived from disaggregating the aggregate of cells.
2. The method of claim 1 , wherein cells are aggregated by the hanging drop method or forced aggregation method.
3. The method of claim 1 , wherein the aggregation of the aggregate is carried out in cell culture.
4. The method of claim 1 , wherein cells in the aggregate and cells derived from the aggregate express one or more of Oct 3/4, telomerase, rex-1, rox-1, nanog, GATA6 and sox-2.
5. The method of claim 1 or 4 , wherein cells in the aggregate and cells derived from the aggregate can differentiate into cell types of all three of the endodermal, ectodermal and mesodermal embryonic lineages.
6. The method of claim 1 , wherein the aggregate contains about 10 cells to about 50,000 cells or more.
7. The method of claim 1 , wherein the aggregate contains about 1000 cells to about 5000 cells.
8. The method of claim 1 or 4 , wherein the non-embryonic stem, non-embryonic germ, non-germ cells are derived from bone marrow.
9. The method of claim 5 , wherein the non-embryonic stem, non-embryonic germ, non-germ cells are derived from bone marrow.
10. The method of claim 8 , wherein the non-embryonic stem, non-germ cells are human cells.
11. The method of claim 9 , wherein the non-embryonic stem, non-germ cells are human cells.
12. The method of claim 1 or 4 further comprising admixing the disaggregated cells with a pharmaceutically-acceptable vehicle for administration to a subject.