Method for producing induced pluripotent stem cells
An object of the present invention is to provide methods for producing iPS cells with low invasivity and high efficiency. The iPS cells can be produced with high efficiency using a method comprising the steps of culturing mononuclear cells derived from peripheral blood for 3 to 14 days in the presence of anti-CD3 antibody, and subjecting the cultured mononuclear cells to dedifferentiation.
1. A method for producing a mammalian induced pluripotent stem cell comprising:
(i) culturing a population consisting essentially of mammalian mononuclear cells isolated from peripheral blood for 3 to 14 days in the presence of anti-CD3 antibody and interleukin 2; and
(ii) subjecting the cultured mononuclear cells to dedifferentiation by introduction of retroviral or Sendai viral vector(s) encoding Oct-4, Sox-2, Klf-4, and c-myc; and
(iii) culturing the cells in (ii) under embryonic stem (ES) cell culture conditions to produce an induced pluripotent stem cell.
2. The method for producing a mammalian induced pluripotent stem cell according to claim 1 , wherein:
the recombinant expression vector is a Sendai virus vector.
3. The method for producing a mammalian induced pluripotent stem cell according to claim 1 , wherein step (iii) comprises culturing the dedifferentiated mononuclear cells for 14 to 25 days in the presence of a growth factor.
4. The method for producing a mammalian induced pluripotent stem cell according to claim 1 , wherein said peripheral blood is isolated from a human.