Screening methods for human embryonic stem cells
This disclosure provides an improved system for culturing human pluripotent stem cells. Traditionally, pluripotent stem cells are cultured on a layer of feeder cells (such as mouse embryonic fibroblasts) to prevent them from differentiating. In the system described here, the role of feeder cells is replaced by components added to the culture environment that support rapid proliferation without differentiation. Effective features are a suitable support structure for the cells, and an effective medium that can be added fresh to the culture without being preconditioned by another cell type. Culturing human embryonic stem cells in fresh medium according to this invention causes the cells to expand surprisingly rapidly, while retaining the ability to differentiate into cells representing all three embryonic germ layers. This new culture system allows for bulk proliferation of pPS cells for commercial production of important products for use in drug screening and human therapy.
1. A method of screening a factor for its effect on a primate pluripotent stem (pPS) cell, comprising:
a) obtaining an undifferentiated pPS cell cultured essentially feeder free on an extracellular matrix in a non-conditioned nutrient media comprising fibroblast growth factor (FGF);
b) contacting the pPS cell with the factor; and
c) detecting any change in the pPS cell.
2. The method of claim 1 , further comprising allowing the pPS cell to differentiate before step b).
3. The method of claim 1 , wherein the factor is a small molecule drug.
4. The method of claim 1 , wherein the factor is a peptide.
5. The method of claim 1 , wherein the factor is a differentiation factor.
6. The method of claim 1 , wherein the detected change is cytotoxicity.
7. The method of claim 1 , wherein the detected change is a change in cellular morphology.
8. The method of claim 1 , wherein the detected change is a change in marker expression.