Induced pluripotent stem cells produced using reprogramming factors and a rho kinase inhibitor or a histone deacetylase inhibitor
Described herein are multipotent stem cells, e.g., human and other mammalian pluripotent stem cells, and related methods.
1. A method for generating a human stem cell, comprising the steps of:
(i) culturing human post-natal cells;
(ii) introducing into the cultured human post-natal cells one or more retroviral vectors encoding Oct 3/4, Sox2, and Klf4; and
(iii) culturing the human post-natal cells from step (ii) in a culture medium suitable for culturing human ES cells, in the presence of an inhibitor of Rho kinase and exogenous FGF-2, on fibroblast feeder cells or on an extracellular matrix;
wherein the culturing is performed under conditions that maintain pluripotency and self-renewal, and wherein exogenous c-Myc is not introduced into the human post-natal cells.
2. The method of claim 1 , wherein the inhibitor of Rho kinase is Y-27632 or HA-1077.
3. A method for generating a human stem cell, comprising the steps of:
(i) culturing human post-natal cells;
(ii) introducing into the cultured human post-natal cells one or more retroviral vectors encoding Oct 3/4, Sox2, and Klf4 and contacting the human post-natal cells with a histone deacetylase inhibitor; and
(iii) culturing the human post-natal cells from step (ii) in a culture medium suitable for culturing human ES cells, in the presence of exogenous FGF-2, on fibroblast feeder cells or on an extracellular matrix;
wherein the culturing is performed under conditions that maintain pluripotency and self-renewal, and wherein exogenous c-Myc is not introduced into the human post-natal cells.