3 replies to this topic

[hrltwc]

Hi

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!!!

[pcrman]

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?

[hrltwc]

by source u mean the target cells that I want to make iPSCs from? If it is, I'm using adult skin fibroblasts...

[biozweistein22]

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.