Methods and compositions for feeder-free pluripotent stem cell media containing human serum
The present invention provides compositions and methods for the culture and maintenance of pluripotent stem cells. More particularly, the present invention provides for compositions and methods for culturing, maintaining, growing and stabilizing primate pluripotent stem cells in a feeder-free defined media further comprising human serum, or a soluble attachment component of the human serum, for promoting cell attachment.
1. A composition comprising primate pluripotent cells, and a medium comprising an effective amount of human serum in a matrix-free culture vessel, wherein the primate pluripotent cells are in contact with the medium in the culture vessel and wherein the serum promotes attachment of the primate pluripotent cells and the cells are attached to the culture vessel.
2. The composition of claim 1 , wherein the medium supports proliferation of the primate pluripotent cell in an undifferentiated state.
3. The composition of claim 1 , wherein the culture vessel does not comprise feeder cells.
4. The composition of claim 1 , wherein the culture vessel is a bioreactor.
5. The composition of claim 1 , wherein the primate pluripotent cells are human embryonic stem cells (hESC).
6. The composition of claim 1 , wherein the human serum is at a concentration of about 0.1% to about 20% of the medium.
7. The composition of claim 1 , wherein the primate pluripotent stem cells continue to proliferate without differentiation for more than 2 serial passages.
8. The composition of claim 1 , wherein the primate pluripotent stem cells are passaged by enzymatic dissociation and replating.