Hi
Is there anyone who have the experience with IPSCs generation? I infected the PBMCs 60 days ago, but i still not find any IPSCs colonies, almost all of them just looks like differentiation. But another line looks good. I use same medium same reprogramming kit, I don't know why the first one not good.Can you please give me some suggestions?
Thank you
Did IPSCs mean inhibitory postsynaptic currents? Why does peripheral blood mononuclear cells which are circulating express synaptic currents? Does peripheral blood cells have "synapses"?
@sheng-nan,
IPSC also means induced pluripotent stem cells
Hello
I agree with all researchers . Also , You can read this study which used frozen PBMCs to generated the iPSC lines in feeder‐free conditions in 16 days. Where used direct reprograming of PBMCs without the need for activation and/or expansion of cell subpopulations within PBMCs.
Article Advanced Feeder-Free Generation of Induced Pluripotent Stem ...
Good Luck
Hi Liyuan,
Just for me to understand, what is different about the 'good line'? Is it simply one line you derived from the same starting population in the same reprogramming? There could very well be differences between clones - that is why it is necessary to pick several clones and continue with the most promising one. Clones also may vary in their proliferation rate, so there are differences one cannot explain.
To come back to the issue, I have also reprogrammed PBMNCs with Sendai virus, and it was quite successful in my case. I have used the reprogramming protocol for infecting with Sendai virus (Thermo Fisher) but for the medium, I used ReproTeSR from the Stem Cell Technologies and followed their protocol. 60 days are definitely long enough to see if the reprogramming is successful (even after 2weeks) and you should be able to establish lines in that amount of time.
Hope this helps, and good luck!