Generation of NHP iPSCs by Sendai virus vector infection

Infection of primary cells was performed with the CytoTune-iPS 2.0 Sendai Reprogramming Kit (Thermo Fisher) at a MOI of 5 using a modified protocol. Briefly, 7 × 10⁵ urine derived cells were incubated in 100 µl of the CytoTune 2.0 SeV mixture containing three vector preparations: polycistronic Klf4–Oct3/4–Sox2, cMyc, and Klf4 for one hour at 37 °C. To control transduction efficiency 3.5 × 10⁵ cells were infected with CytoTune-EmGFP SeV. Infected cells were seeded on Geltrex (Thermo Fisher) coated 12-well-plates, routinely 10 × 10³ and 25 × 10³ cells per well. Medium was replaced with fresh Renal epithelial and mesenchymal cell proliferation medium RE/MC (ATCC) every second day. On day 5, medium was changed to mTeSR1 (Stemcell Technologies), with subsequent medium changes every second day. After single colony picking, cells were cultured in StemFit (Ajinomoto) supplemented with 100 ng/ml recombinant human basic FGF (Peprotech), 100 U/ml Penicillin and 100 μg/ml Streptomycin (Thermo Fisher).