Compositions and methods for deriving or culturing pluripotent cells
OwnerYaqub Hanna, Rudolf Jaenischinventors
Life of this patent
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Abstract
The invention provides compositions and methods useful for deriving or culturing vertebrate ES cells. Certain inventive methods comprise deriving or culturing vertebrate ES cells using medium that comprises a compound that replaces Klf4 or c-Myc in generating iPS cells. The invention provides NOD ES cells and methods of deriving or culturing them.
Claims27
A non-obese diabetic (NOD) embryonic stem (ES) cell.
-18. (canceled)
A method of producing a chimeric mouse that comprises cells derived from a NOD mouse, the method comprising steps of:
- (a) providing a NOD ES cell;
- (b) introducing the NOD ES cell into a mouse blastocyst;
- (c) transferring the blastocyst to a pseudopregnant female mouse;
- and
- (d) maintaining the female mouse under conditions suitable for production of live offspring.
-22. (canceled)
A vertebrate cell isolated from the ICM of a vertebrate blastocyst or from the epiblast of a vertebrate embryo, wherein the vertebrate cell has been caused to express Klf4 or c-Myc.
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A composition comprising:
- (a) a vertebrate cell isolated from the ICM of a vertebrate blastocyst or isolated from the epiblast of a vertebrate embryo;
- and (b) a compound that replaces Klf4 or c-Myc expression in generating iPS cells.
-45. (canceled)
A method of deriving a vertebrate ES cell, the method comprising:
- (a) providing a vertebrate cell isolated from the ICM of a vertebrate blastocyst or isolated from the epiblast of a vertebrate embryo;
- (b) causing the cell to persistently express Klf4 or c-Myc or culturing the cell in medium that contains a compound that replaces Klf4 or c-Myc expression in generating iPS cells;
- and (c) maintaining the cell in culture under conditions suitable and a time sufficient to produce a vertebrate ES cell.
-57. (canceled)
A method of culturing a vertebrate ES cell, the method comprising:
- (a) providing a vertebrate ES cell;
- and (b) culturing the cell in medium that contains a compound that replaces Klf4 or c-Myc expression in generating iPS cells.
-66. (canceled)
A composition comprising an induced pluripotent stem (iPS) cell and a compound that replaces Klf4 or c-Myc expression in generating iPS cells, wherein said iPS cell is unstable in the absence of the compound.
-90. (canceled)
A method of deriving an iPS cell from a somatic cell of a NOD mouse comprising steps of:
- (a) providing a somatic cell obtained from a NOD mouse;
- (b) causing the cell to express a set of reprogramming factors or contacting the cell with a set of reprogramming compounds sufficient to induce the cell to become reprogrammed to a pluripotent state;
- and
- (c) causing the cell to persistently express Klf4 or c-Myc or culturing the cell in medium that contains a compound that replaces Klf4 or c-Myc expression in generating iPS cells.
A method of producing a chimeric mouse that comprises cells derived from a NOD mouse, the method comprising steps of:
- (a) providing an iPS cell derived from a somatic cell of a NOD mouse;
- (b) introducing the iPS cell into a mouse blastocyst;
- (c) transferring the blastocyst to a pseudopregnant female mouse;
- and
- (d) maintaining the female mouse under conditions suitable for production of live offspring.
(canceled)
A method of modifying the pluripotency state of a vertebrate cell to a more naïve state, the method comprising:
- (a) providing a pluripotent vertebrate cell;
- (b) causing the cell to express Klf4 and/or Klf2;
- (c) culturing the cell in the presence of a GSK3 inhibitor, a mitogen-activated protein kinase pathway inhibitor, and an activator of the Stat3 pathway;
- and (d) maintaining the cell in culture under conditions suitable and a time sufficient to produce a vertebrate cell having a resulting pluripotency state which is more naïve than the pluripotency state of the vertebrate cell of step (a).
-130. (canceled)
A pluripotent vertebrate cell which has been caused to upregulate expression of KLf4 and/or Klf2 and in which the GSK3 pathway has been inhibited, the mitogen-activated protein kinase pathway has been inhibited, and the Stat3 pathway has been activated.
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A method of culturing a vertebrate cell, the method comprising:
- (a) providing a pluripotent vertebrate cell;
- (b) causing the cell to upregulate expression of Klf4 and/or Klf2;
- (c) culturing the cell in the presence of a GSK3 inhibitor, a mitogen-activated protein kinase pathway inhibitor, and an activator of the Stat3 pathway.
-157. (canceled)
Dependencies are inferred from the claim text.