Hello friends,
Does anybody here work with small molecules to induce pluripotency in human somatic cells and can ask a few technical questions:
1. Which media do you use and do you switch to other media after a certain period of time?
2. Do you add the chemicals to the medium every time you change the medium or you add then only once and that's it?
3. Where do you keep the stock solutions (+4 or -20)?
4. Would love to get a protocol if possible
Thanks a lot in advance,
Anna
What cell type are you looking to reprogram? Generally human fibroblasts have the highest efficiency for reprogramming. There are several kits available from Stem Cell Technologies and Reprocell. I have reprogrammed fibroblasts and AD-MSCs using RNA-based kits that include all the micro-RNAs for pluripotency (Oct4, Soz2, Klfq4, cMyc, Nanog and Ling28). The kit is stored at -80. It involves a series of 4 transfections using Rnaimax from Thermofisher. Colonies are visible after 10-14 days. We use Nutristem medium from Reprocell.
Heather Wanczyk Thanks for the reply! I’m Iooking to reprogram human dermal fibroblasts but not with genes such as Oct4, Sox2 etc. but with chemical compounds that substitute these factors
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