METHOD FOR CULTURING STEM CELLS
First Claim
1. A method for culturing embryonic stem cells to obtain a population of embryoid bodies, the method comprising the steps of:
- harvesting substantially undifferentiated embryonic stem cells from a dimension-constrained microwell; and
culturing the harvested embryonic stem cells under differentiating conditions to form the population of embryoid bodies.
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Accused Products
Abstract
A three-dimensional microwell system that supports long term embryonic stem cell (ESCs) culture and formation of homogeneous embryoid bodies (EBs) is described. Microwell-cultured ESCs remain viable and undifferentiated for several weeks in culture and maintain undifferentiated replication when passaged to Matrigel®-coated, tissue culture-treated polystyrene dishes. Microwell-cultured ESCs maintain pluripotency, differentiating to each of the three embryonic germ layers. ESC aggregates released from microwells can be passaged for undifferentiated replication or differentiated to monodisperse EBs. The ability to constrain ESC growth in three dimensions advantageously provides for more efficient, reproducible culture of undifferentiated cells, high-throughput screening, and the ability to direct ESC differentiation by generating monodisperse EBs of a desired size and shape.
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Citations
17 Claims
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1. A method for culturing embryonic stem cells to obtain a population of embryoid bodies, the method comprising the steps of:
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harvesting substantially undifferentiated embryonic stem cells from a dimension-constrained microwell; and
culturing the harvested embryonic stem cells under differentiating conditions to form the population of embryoid bodies. - View Dependent Claims (2, 3, 4, 5, 6, 7)
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- 8. An array comprising a substrate having a polymer coating thereupon, the coating defining a microwell on an upon surface thereof, the microwell having side walls and a bottom and having a cell-attractive portion and a cell-repulsing portion.
- 12. Undifferentiated human embryonic stem cell colonies having a substantially uniform size and shape.
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14. An embryonic stem cell-derived embryoid body having a substantially uniform size and shape.
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15. A method of cryopreserving embryonic stem cells, comprising the step of:
freezing microwell-cultured, embryonic stem cells in microwells. - View Dependent Claims (16)
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17. A method for culturing embryonic stem cells, the method comprising the step of:
culturing substantially undifferentiated embryonic stem cells in a dimension-constrained microwell without subculture for at least about three weeks in a medium that does not promote differentiation, wherein greater than about 90% of the embryonic stem cells remain undifferentiated after about three weeks.
Specification