西亚试剂：Mononuclear (MNC) blood cells were isolated from several do

Mononuclear (MNC) blood cells were isolated from several donors by Ficoll-Hypaque density gradient centrifugation (Ferrante and Thong, 1980,Vissers et al., 1988). Samples were frozen and thawed days to several months after freezing and expanded in IL-7 or in G-CSF, GM-CSF, IL-6, and IL-3 for 5 days. In our initial experiments we used the FUW-M2rtTA and the individual doxycycline-inducible lentiviruses encoding Oct4, Sox2, c-Myc, or Klf4 (Brambrink et al., 2008). However, in ten independent experiments we were not able to reprogram peripheral blood cells with this system. One possibility for failure to obtain iPSCs is that peripheral blood cells are difficult to infect, reducing the probability of obtaining cells carrying the four factors as well as the FUW-M2rtTA construct. Also, we find that the efficiency of blood reprogramming (0.001%–0.0002%) is approximately 10–50 times lower than that of human fibroblast reprogramming.

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