Non-static suspension culture of cell aggregates
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, endodermal, and mesodermal embryonic germ layers, e.g., stem cells.
1. A method for making a cell culture composition comprising aggregates of stem cells that are not embryonic stem cells, embryonic germ cells, or germ cells, wherein the aggregates are in non-static cell culture and wherein the cell density ranges from about 5×10 4 cells/ml to about 10 8 cells/ml, the method comprising inoculating a cell culture with stem cell aggregates and expanding the inoculated aggregates so that expansion produces a cell density of about 5×10 4 cells/ml to about 10 8 cells/ml.
2. The method of claim 1 wherein the cells that are not embryonic stem cells, embryonic germ cells, or germ cells can differentiate into cell types of at least two of the endodermal, ectodermal and mesodermal embryonic lineages.
3. The method of claim 1 wherein the non-embryonic stem cells, non-embryonic germ cells, non-germ cells express oct4, are not transformed, and have a normal karyotype.
4. The method of claim 1 wherein the non-embryonic stem cells, non-embryonic germ cells, non-germ cells are not transformed and have a normal karyotype, and wherein, prior to aggregation, the cells have undergone about 40 cell doublings in culture.
5. The method of claim 1 wherein the non-embryonic stem cells, non-embryonic germ cells, non-germ cells express telomerase, are not transformed, and have a normal karyotype.
6. The method of claim 1 wherein the non-embryonic stem cells, non-embryonic germ cells, non-germ cells are positive for one or more of oct3/4, telomerase, rex-1, rox-1, nanog, GATA6 and sox-2.
7. The method of any of claims 1 and 2 - 6 wherein expansion is non-adherent.
8. The method of any of claims 1 and 2 - 6 wherein the non-embryonic stem cells, non-embryonic germ cells, non-germ cells are derived from bone marrow.
9. The method of claim 7 wherein the non-embryonic stem cells, non-embryonic germ cells, non-germ cells are derived from bone marrow.