Establishment of iPSCs from Urine Cells

Establishment of iPSCs was performed as reported before^{18 (link)}. 0.5 × 10⁶ primary urine cells were reprogrammed using the Amaxa Basic Nucleofector Kit (Lonza) according to the protocol of the manufacturer. 1 μg of each plasmid pCXLE-hOCT3/4-shp53-F (Addgene #27077), pCXLE-hSK (Addgene #27078), and pCXLE-hUL (Addgene #27080) was used. After nucleofection, the cells were transferred to a Matrigel (Corning) coated 6 well. After 24 h, the UC medium was replaced by mTeSR-1 (Stemcell Technologies), supplemented with 1% PenStrep, and changed every second day. After about two weeks the emerged iPSC colonies were picked and transferred to a new Matrigel-precoated 6 well plate. The mTeSR-1 cell medium (was replaced on a daily basis thereafter and the iPSC colonies were subcultured every seven days using 1 U/ml Dispase (Stemcell Technologies). During cultivation, colonies with atypical morphology were removed by scraping. HepaRG cells served as biliary reference cell line and were maintained in RPMI/10% FCS.