Transcriptional profiling of all KMT2B-WT, HT, and Int HPCs. KMT2B-HT and Int iPSCs

HBV-KMT2B integrated human induced pluripotent stem cell-derived hepatic progenitor cells (KMT2B-Int HPCs) vs heterozygous mutated KMT2B human induced pluripotent stem cell-derived hepatic progenitor cells (KMT2B-HT HPCs) vs original human induced pluripotent stem cell-derived hepatic progenitor cells (KMT2B-WT HPCs) Transcriptional profiling of all KMT2B-WT, HT, and Int HPCs. KMT2B-HT and Int iPSCs were generated by gene editing using CRISPR/Cas9 and Cre recombinase technologies. iPSCs were differentiated into a hepatic lineage. On Day14 of hepatic differentiation, CD13 high and C133 high hepatic cells were sorted by fluorescence-activated cell sorter (FACS) on feeder mouse embryonic fibroblasts (MEFs). HPCs forms colony on MEFs and were stably cultured with maintaining their hepatic phenotype. Transcriptional profiling of all KMT2B-WT, HT, KO, Int iPSCs. As the KMT2B-Wand T iPSCs, P11025 iPSCs were used, which were reprogrammed by the episomal vector from normal skin. KMT2B-HT, KO, and Int iPSCs were generated by gene editing using CRISPR/Cas9 and Cre recombinase technologies. KMT2B-HT iPSCs were used as control cell lines of KMT2B-Int iPSCs because KMT2B Int iPSCs were heterozygous KMT2B due to HBV integration. DNA sequences as results of gene editing such as integration of HBV sequences were determined by direct sequencing. Knockout of KMT2B also was confirmed by immunoblotting. All iPSCs were differentiated into hepatic lineages. At day14 of hepatic differentiation, CD13 high and CD133 high hepatic cells were sorted on MEFs After 2week cultivation, colonies of HPCs were cultured and then passaged. These HPCs maintained hepatic phenotype and proliferative potential. All HPCs were passaged on MEFs and cultured for about 14 days, and the HPCs were harvested and analyzed.