Method For Producing Induced Pluripotent Stem Cells

Abstract

Provided is a method of generating iPS cells. Specifically, provided is a method of generating iPS cells having a rearranged γδ-TCR gene. Also provided is a cell population including the generated iPS cells. The method includes stimulating collected blood cells with IL-2 and a bisphosphonate, and then introducing cell reprogramming factors through use of a Sendai virus (SeV) vector. According to the method of the present invention, iPS cells having a rearranged γδ-TCR gene can be effectively generated. In particular, the method may be free of a step of treating the blood cells with an antibody before the step of stimulating blood cells with any one kind or a plurality of kinds of interleukins selected from IL-2, IL-15, and IL-23, and a bisphosphonate. In addition, iPS cells generated by the method of the present invention can be differentiated into desired cells by differentiation induction treatment.

1 . A method of generating iPS cells, comprising the following steps 1) to 3):
 1) stimulating collected blood cells with any one kind or a plurality of kinds of interleukins selected from IL-2, IL-15, and IL-23, and a bisphosphonate;   2) introducing at least four kinds of genes capable of expressing cell reprogramming factors into the blood cells through use of a Sendai virus vector; and   3) culturing the cells having introduced therein the genes.   
     
     
         2 . The method of generating iPS cells according to  claim 1 , wherein the cell reprogramming factors comprise OCT3/4, SOX2, KLF4, and c-MYC. 
     
     
         3 . The method of generating iPS cells according to  claim 1 , wherein the interleukins comprise IL-2. 
     
     
         4 . The method of generating iPS cells according to  claim 1 , wherein the bisphosphonate comprises one kind or a plurality of kinds selected from zoledronic acid, pamidronic acid, alendronic acid, risedronic acid, ibandronic acid, incadronic acid, etidronic acid, minodronic acid, salts thereof, and hydrates thereof. 
     
     
         5 . The method of generating iPS cells according to  claim 1 , wherein the blood cells comprise peripheral blood mononuclear cells. 
     
     
         6 . The method of generating iPS cells according to  claim 1 , wherein the blood cells comprise cells of human origin. 
     
     
         7 . The method of generating iPS cells according to  claim 1 , wherein the method is free of a step of treating the collected blood cells with an antibody before the steps 1) to 3). 
     
     
         8 . The method of generating iPS cells according to  claim 1 , wherein the iPS cells comprise iPS cells having a rearranged γδ-TCR gene. 
     
     
         9 . A cell population, comprising iPS cells generated by the method of generating iPS cells according to  claim 1 . 
     
     
         10 . Blood progenitor cells, which are obtained by inducing differentiation of iPS cells generated by the method of generating iPS cells according to  claim 1 .