Induction of pluripotent cells
First Claim
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1. A composition comprising:
- an activin receptor-like kinase 5 (ALK5) inhibitor;
a MAP/ERK kinase (MEK) inhibitor; and
a Rho kinase (ROCK) inhibitor, wherein the ALK5 inhibitor, the MEK inhibitor, and the ROCK inhibitor are present in amounts effective to increase efficiency of reprogramming a non-pluripotent mammalian cell to a pluripotent cell, as compared to non-pluripotent mammalian cells treated with the ALK5 inhibitor and the MEK inhibitor without the ROCK inhibitor.
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Abstract
The slow kinetics and low efficiency of reprogramming methods to generate human induced pluripotent stem cells (iPSCs) impose major limitations on their utility in biomedical applications. Here we describe a chemical approach that dramatically improves (>200 fold) the efficiency of iPSC generation from human fibroblasts, within seven days of treatment. This will provide a basis for developing safer, more efficient, non-viral methods for reprogramming human somatic cells.
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Citations
18 Claims
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1. A composition comprising:
- an activin receptor-like kinase 5 (ALK5) inhibitor;
a MAP/ERK kinase (MEK) inhibitor; and
a Rho kinase (ROCK) inhibitor, wherein the ALK5 inhibitor, the MEK inhibitor, and the ROCK inhibitor are present in amounts effective to increase efficiency of reprogramming a non-pluripotent mammalian cell to a pluripotent cell, as compared to non-pluripotent mammalian cells treated with the ALK5 inhibitor and the MEK inhibitor without the ROCK inhibitor. - View Dependent Claims (2, 3, 4, 5, 6, 17)
- an activin receptor-like kinase 5 (ALK5) inhibitor;
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7. A mixture comprising:
- one or more mammalian cells;
an ALK5 inhibitor;
a MEK inhibitor; and
a ROCK inhibitor, wherein the ALK5 inhibitor, the MEK inhibitor, and the ROCK inhibitor are present in amounts effective to increase efficiency of reprogramming a non-pluripotent mammalian cell to a pluripotent cell, as compared to non-pluripotent mammalian cells treated with the ALK5 inhibitor and the MEK inhibitor without the ROCK inhibitor. - View Dependent Claims (8, 9, 10, 18)
- one or more mammalian cells;
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11. A method of inducing non-pluripotent mammalian cells into induced pluripotent stem cells, comprising:
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(a) introducing Oct-3/4, Klf4, Sox2, and optionally c-Myc into the non-pluripotent cells; and (b) contacting the non-pluripotent cells with an ALK5 inhibitor, a MEK inhibitor, and a ROCK inhibitor, wherein the ALK5 inhibitor, the MEK inhibitor, and the ROCK inhibitor increase efficiency of inducing the non-pluripotent mammalian cells into induced pluripotent stem cells, as compared to non-pluripotent mammalian cells treated with the ALK5 inhibitor and the MEK inhibitor without the ROCK inhibitor. - View Dependent Claims (12, 13, 14, 15, 16)
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Specification