Stem Cell Aggregates and Methods for Making and Using

Abstract

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 - 84 . (canceled) 
     
     
         85 . A method comprising disaggregating cells in an aggregate, wherein the cells in the aggregate that are disaggregated are homogenous, the aggregate having been formed by exposing cells that are not embryonic stem cells, embryonic germ cells, or germ cells and express telomerase, are not tumorigenic, and have a normal karyotype, 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. 
     
     
         86 . The method of  claim 85 , wherein cells in the aggregate and cells derived from the aggregate express one or more of Oct 3/4, rex-1, rox-1, nanog, GATA6 and sox-2. 
     
     
         87 . The method of  claim 85  or  86 , wherein cells in the aggregate and cells derived from the aggregate can differentiate into cell types of at least two of the endodermal, ectodermal, mesodermal embryonic lineages. 
     
     
         88 . The method of  claim 87 , 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. 
     
     
         89 . The method of  claim 85 , wherein the non-embryonic stem, non-germ cells are human cells. 
     
     
         90 . The method of  claim 86 , wherein the non-embryonic stem, non-germ cells are human cells. 
     
     
         91 . The method of  claim 89 , wherein the non-embryonic stem, non-embryonic germ, non-germ cells are derived from bone marrow. 
     
     
         92 . The method of  claim 90 , wherein the non-embryonic stem, non-embryonic germ, non-germ cells are derived from bone marrow. 
     
     
         93 . The method of  claim 85 , wherein cells are aggregated by the hanging drop method or forced aggregation method. 
     
     
         94 . The method of  claim 85 , wherein the aggregation of the aggregate is carried out in cell culture. 
     
     
         95 . The method of  claim 85 , wherein the aggregate contains about 10 cells to about 50,000 cells or more. 
     
     
         96 . The method of  claim 85 , wherein the aggregate contains about 1000 cells to about 5000 cells. 
     
     
         97 . The method of  claim 85  or  86  further comprising admixing the disaggregated cells with a pharmaceutically-acceptable vehicle for administration to a subject.