Induction of pluripotent cells
AssigneeThe Scripps Research Institute
InventorsTongxiang Lin, Sheng Ding
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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.
Claims21
A composition comprising:
- (i) a Rho kinase (ROCK) inhibitor;
- (ii) a glycogen synthase kinase 3 (GSK3) inhibitor;
- and
- (iii) an ALK5 inhibitor or a MAP/ERK kinase (MEK) inhibitor;
- wherein the ROCK inhibitor has the structure:
- wherein,
- L2 is unsubstituted C1-C10 alkylene;
- y is an integer from 0 to 1;
- z is an integer from 0 to 1;
- X is —N═, or —CH═;
- R1 is hydrogen or unsubstituted C1-C6 alkyl;
- R3 is —OR18, or substituted or unsubstituted C1-6 alkyl, wherein the substituted C1-6 alkyl is substituted with halogen;
- R4 is —OR18;
- and
- R18 is hydrogen or unsubstituted C1-6 alkyl;
- or a pharmaceutically acceptable salt thereof.
A mixture comprising:
- one or more mammalian cells;
- a Rho kinase (ROCK) inhibitor;
- a GSK3 inhibitor;
- and
- an ALK5 inhibitor or a MAP/ERK kinase (MEK) inhibitor;
- wherein ROCK inhibitor has the structure:
- wherein
- L2 is unsubstituted C1-C10 alkylene;
- y is an integer from 0 to 1;
- z is an integer from 0 to 1;
- X is —N═, or —CH═;
- R1 is hydrogen or unsubstituted C1-C6 alkyl;
- R3 is —OR18, or substituted or unsubstituted C1-6 alkyl, wherein the substituted C1-6 alkyl is substituted with halogen;
- R4 is —OR18, and
- R18 is hydrogen or unsubstituted C1-6 alkyl;
- or a pharmaceutically acceptable salt thereof.
A method of inducing non-pluripotent mammalian cells into induced pluripotent stem cells in vitro, comprising:
- (a) introducing an exogenous transcription factor comprising Oct-3/4, and optionally one or more of Klf, Sox2, and c-Myc;
- and
- (b) contacting the non-pluripotent cells with a ROCK inhibitor under conditions sufficient for inducing non-pluripotent mammalian cells into induced pluripotent stem cells;
- wherein ROCK inhibitor has the structure:
- wherein,
- L2 is unsubstituted C1-C10 alkylene;
- y is an integer from 0 to 1;
- z is an integer from 0 to 1;
- X is —N═, or —CH═;
- R1 is hydrogen or unsubstituted C1-C6 alkyl;
- R3 is —OR18, or substituted or unsubstituted C1-6 alkyl, wherein the substituted C1-6 alkyl is substituted with halogen;
- R4 is —OR18;
- and
- R18 is hydrogen or unsubstituted C1-6 alkyl;
- or a pharmaceutically acceptable salt thereof.
Dependencies are inferred from the claim text.