We standardly generate transgene-free iPSCs by Sendai virus vectors, which provides a safer and non-integrating alternative to traditional reprogramming methods. iPSC lines are only issued after we proved RNA reprogramming vectors are cleared from iPSCs and after mycoplasma testing.
Our standard iPSC generation package includes 3 different iPSC lines per individual (“clones”). More clones or use of other reprogramming methods can be requested.
It is essential to confirm pluripotency in the newly generated iPSC lines. Unless requested differently, our standard iPSC generation package includes immunocytochemistry for pluripotency markers (OCT3/4, SSEA4, TRA-1-60 and TRA-1-81) and spontaneous (“embryoid body”) differentiation into the three germ layers. We provide and recommend G-banding karyotyping to determine chromosome integrity.
We facilitate registration of iPSC lines at the European Human Pluripotent Stem Cell Registry (www.hpscreg.eu). A hPSCreg® certificate evidences for ethical provenance and scientific evidence for pluripotency using standardized criteria. It is possible to opt-out from registration.
Time line and costs
It takes 6 months to obtain fully characterized iPSC lines, but this is subjected to preferences and the current availability of iPSC services. Standard full service costs per donor is 2990 euro. For intake and special requests contact: iPSC@amsterdamumc.nl