Micro-carrier culture system for rapid expansion of human embryonic stem cells
First Claim
1. A method of expanding primate pluripotent stem cells (pPS cell) in an undifferentiated state comprising a) attaching a pPS cell to a microcarrier comprising a matrix and b) providing the pPS cells with a nutrient media and c) expanding 2 fold the pPS cells in an undifferentiated state.
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Abstract
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.
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Citations
24 Claims
- 1. A method of expanding primate pluripotent stem cells (pPS cell) in an undifferentiated state comprising a) attaching a pPS cell to a microcarrier comprising a matrix and b) providing the pPS cells with a nutrient media and c) expanding 2 fold the pPS cells in an undifferentiated state.
- 13. A method of expanding cells expressing the markers Oct 4, stage specific antigen 4 (SSEA4), Tra-1-60 and Tra-1-81 in an undifferentiated state comprising a) attaching the cells expressing the markers Oct 4, stage specific antigen 4 (SSEA4), Tra-1-60 and Tra-1-81 to a microcarrier comprising a matrix and b) providing the cells expressing the markers Oct 4, stage specific antigen 4 (SSEA4), Tra-1-60 and Tra-1-81 with a nutrient media and c) expanding 2 fold the cells expressing, the markers Oct 4, stage specific antigen 4 (SSEA4), Tra-1-60 and Tra-1-81 in an undifferentiated state.
Specification