Bile as a non-invasive source of cholangiocyte organoids for develping patient-specific disease modelling and personalized regenerative medicine

F.J.M. Roos, M.M.A. Verstegen, J.W. Poley, M. Bruno, G.W.M. Tetteroo, J.N.M. IJzermans, L.J.W. van der Laan

Wednesday 13 march 2019

16:50 - 17:00h at Tropentheater

Categories: Best abstracts, Parallelsessie

Parallel session: Parallelsessie VIII – Best abstracts II

Background: Bile duct related diseases are the leading cause for pediatric liver transplantation and adult re-transplantation of a liver graft. Studying biliary diseases has long-term been hampered by the inability to culture bile duct lining cholangiocytes long-term. Recently was shown that Extra-hepatic Cholangiocyte Organoids (ECOs) that are derived from extra-hepatic bile duct (EHBD) tissue can be long-term expanded in culture. However, disease modeling or personalized regenerative medicine applications are limited since highly invasive bile duct biopsies are required to obtain these ECOs from individual patients. Therefore the aim of the current study is to investigate whether ECOs can be cultured from less invasively acquired bile fluid.

Methods: Bile-derived cholangiocyte organoids (BCOs) were cultured, according to the previous published protocol and collected from gallbladder bile obtained from donor livers for transplantation and from bile obtained by endoscopic retrograde cholangiopancreatography (ERCP) or percutaneous transhepatic cholangiography drain (PTCD) in patients. In addition, ECOs were initiated from three different patients and compared to BCOs on the genetic level (qrt-PCR), protein level (either immunohistochemistry, immunofluorescence or Western blotting) and functional level by testing the cholangiocyte specific transporter channels (ussing chamber and transport assay).

Results: Cultures were initiated from 1 ml of bile obtained from all different sources. Bile-derived cholangiocyte organoids could be effectively (8/9 attempts) expanded from all sources of bile from patients with a variety of diseases (primary sclerosing cholangitis, cholangiocarcinoma, bile stones and biliary stenosis after liver transplantation). BCOs expressed similar cholangiocyte markers on gene and protein level as tissue-derived ECOs and both lacked either stem cell- or hepatocyte markers. Furthermore, these cells expressed and responded similarly to stimulation and inhibition of different cholangiocyte ion-channels. Interestingly, cholangiocyte-organoids from a patient with cystic fibrosis (CF) clearly lacked CFTR channel activity, showing that cholangiocyte-organoids can be used as a disease model to study biliary diseases.

Conclusions: Our study showed that bile provides a novel minimally-invasive source of patient-specific cholangiocyte organoids. This creates new opportunities to study autologous bile duct regeneration and develop patient-specific disease models.