Hi,
I am new to this field. I want good links and protocol related to iPSC preparation. Can we prepare iPSC from lung tissue? Do we need immortalization step during this process.
Hello Deepak,
There are lots of kits available in the market now and I think the most popular these days which we use in USA are Sendai Virus Reprogramming kits. You can easily get a pamphlet with detailed protocol on internet for that. Best and easiest cells till date have been embryonic fibroblasts and most people do with them but many people try adult fibroblasts also but efficiency is low into them. Now people are also reprogramming endothelial cells and many other type of cells. You can obviously prepare iPSCs from lung tissue but I am not sure about the reprogramming efficiency you are gonna get from them. The kits provide with Tra-1 antibody where you can select and pick the positive colonies. Then you can grow these colonies and upscale iPSCs.
Hope it helps,
Ajay
You're not gonna need to immortalize them, TERT will be expressed endogenously upon acquisition of pluripotency. I'd maybe start with Sendai virus as a vector and for reprogramming and culture of iPSC vitronectin and E8 medium. The only thing is, buying reprogramming vectors from commercial sources can be expensive, so you can also look into episomal reprogramming, get a few plasmids from Addgene and have plenty of vector material to do lots of experiments with.
- Badges
- Science topic