Maintenance of embryonic stem cells by the GSK-3 inhibitor 6-bromoindirubin-3'-oxime
First Claim
1. A method of maintaining the undifferentiated state of an embryonic stem cell, said method comprising contacting the stem cell in vitro with a molecule that antagonizes GSK-3 activity such that the cell divides but does not differentiate, wherein said contacting is in the absence of a feeder layer.
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Abstract
The present invention relates to methods for maintaining the undifferentiated state of embryonic stem cells without the use of a feeder layer by activating the Wnt signal transduction pathway or by inhibiting glycogen synthase kinase-3 activity by contacting the cell with, inter alia, 6-bromoindirubin-3′-oxime. The present invention also relates to embryonic stem cell lines and cells derived therefrom that have been isolated and cultured in the absence of a feeder layer.
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Citations
31 Claims
- 1. A method of maintaining the undifferentiated state of an embryonic stem cell, said method comprising contacting the stem cell in vitro with a molecule that antagonizes GSK-3 activity such that the cell divides but does not differentiate, wherein said contacting is in the absence of a feeder layer.
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6. A method of maintaining the undifferentiated state of an embryonic stem cell, said method comprising contacting said stem cell with 6-bromoindirubin-3′
- -oxime such that the cells divides but does not differentiate.
- View Dependent Claims (7, 8, 9)
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10. An isolated embryonic stem cell in contact with 6-bromoindirubin-3′
- -oxime.
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11. An embryonic stem cell that is the progeny of a second embryonic stem cell that was previously contacted with 6-bromoindirubin-3′
- -oxime.
- View Dependent Claims (12)
- 13. An embryonic stem cell line produced by the process comprising isolating embryonic stem cells from an embryo and culturing the isolated embryonic stem cells in the presence of a molecule that antagonizes GSK-3 activity such that the isolated embryonic stem cells divide but do not differentiate, wherein said isolating and culturing is in the absence of a feeder layer.
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17. An embryonic stem cell line produced by the process comprising isolating embryonic stem cells from an embryo and culturing the isolated cells in the presence of 6-bromoindirubin-3′
- -oxime such that the isolated embryonic stem cells divide but do not differentiate.
- View Dependent Claims (18, 19)
- 20. A method of obtaining an embryonic stem cell line comprising isolating embryonic stem cells from an embryo and culturing the isolated embryonic stem cells in the presence of a molecule that antagonizes GSK-3 activity such that the isolated embryonic stem cells divide but do not differentiate, wherein said isolating and culturing is in the absence of a feeder layer.
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24. A method of obtaining an embryonic stem cell line comprising isolating embryonic stem cells from an embryo and culturing the isolated embryonic stem cells in the presence of 6-bromoindirubin-3′
- -oxime such that the embryonic stem cells divide but do not differentiate.
- View Dependent Claims (25, 26)
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27. A human embryonic cell line, which cell line has not been in contact with a feeder layer and has not been in contact with an exogenous cell extract.
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28. A human embryonic cell line, which cell line has not been in contact with a feeder layer and has not been in contact with a recombinant human protein.
Specification