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 cell culture composition comprising aggregates of stem cells in non-static cell culture wherein the cell density ranges from about 5×10 4 cells/ml to about 10 8 cells/ml, wherein the number of cells per aggregate is about 50,000 cells and the average diameter of each aggregate is about 1 mm.
2. A method for making the cell culture composition of claim 1 , 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, wherein the number of cells per aggregate is about 50,000 cells and the average diameter of each aggregate is about 1 mm.
3. The cell culture composition of claim 1 wherein the aggregate of cells comprises 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 and mesodermal embryonic lineages.
4. The cell culture composition of claim 1 wherein the aggregate of cells comprises cells that are isolated expanded non-embryonic stem, non-germ cells, the cells having undergone about 10-40 cell doublings in culture, wherein the cells express oct4, are not transformed, and have a normal karyotype.
5. The cell culture composition of claim 1 wherein the aggregate of cells comprises cells that are isolated expanded non-embryonic stem, non-germ cells that are obtained by culture of non-embryonic, non-germ tissue, the cells having undergone about 40 cell doublings in culture, wherein the cells are not transformed and have a normal karyotype.
6. The cell culture composition of claim 1 wherein the aggregate of cells comprises cells that are isolated expanded non-embryonic stem, non-germ cells, the cells having undergone about 10-40 cell doublings in culture, wherein the cells express telomerase, are not transformed, and have a normal karyotype.
7. The cell culture composition of claim 1 wherein the aggregate of cells comprises cells that are isolated expanded non-embryonic stem, non-germ cells that can differentiate into at least one cell type of at least two of the endodermal, ectodermal, and mesodermal embryonic lineages, said cells having undergone about 10-40 cell doublings in culture.
8. The cell culture composition of claim 1 wherein the aggregate of cells comprises cells that are positive for one or more of oct3/4, telomerase, rex-1, rox-1, nanog, GATA6 and sox-2.