GSM5708893: RNA_seq_WT_pregnant-dpc8_biliary epitheial cells_ex3; Mus musculus; RNA-Seq
Mice were laparotomized under isoflurane inhalation anesthesia, and the portal vein was cannulated and perfused with liver perfusion medium. Then, the liver was perfused with liver digestion medium. To isolate BECs, remaining undigested liver pieces were further treated with collagenase, DNase I, and Dispase. The cell suspension was filtered with 70 micrometer cell strainer, then the filtrates were centrifugation at 50g to remove the parenchymal fraction. The non-parenchymal fraction in the supernatant was stained with anti-FcR antibody (rat, 1:100, Bio Legend, 101302), followed by staining with the fluorescence-conjugated antibodies for 1 h. After wash with PBS plus 1 % BSA, the cells were sorted by FACS. RNA was extracted from the FACS-sorted BECs by using an RNeasy Micro Kit (Qiagen). 50 ng Total RNA was used to generate RNA-seq libraries using KAPA RNA HyperPrep Kit with RiboErase (Roche).