iPSC - (May/16/2011 )
I'm trying to make iPSCs using the lentiviral method, but after 10 times of trying, I cant get anything.. not a single colony appears. anyone with experience on what to look out for?
Thanks a lot!!!
the lentiviral platform and Thomson's recipe are less effective than retroviral Yamanaka's recipe. So you may want to switch to retrovirus. By the way, what source cells are you using?
by source u mean the target cells that I want to make iPSCs from? If it is, I'm using adult skin fibroblasts...
First of all , you cannot go switching protocols especially in the iPSCs , each one has a different target. For example , if you want to create integration free iPSCs then you can go with the proteins , mRNA , plasmids , episomal vectors... if you do not care about integration then you can go with yamanaka's ''recipie'', or the lentiviral system which offer greater efficiency.
I have personally used the lentiviral system and it worked fine.
If you can post your protocol , I'll be happy to pinpoint our differences.