Protocol for inducing branching morphogenesis in human cholangiocyte and cholangiocarcinoma organoids

Kimberley Ober; Floris J M Roos; Gilles S van Tienderen; Kübra Köten; Annelot Klaassen; Wunan Mi; Luc J W van der Laan; Monique M A Verstegen

doi:10.1016/j.xpro.2023.102431

Protocol for inducing branching morphogenesis in human cholangiocyte and cholangiocarcinoma organoids

STAR Protoc. 2023 Sep 15;4(3):102431. doi: 10.1016/j.xpro.2023.102431. Epub 2023 Jul 9.

Authors

Kimberley Ober¹, Floris J M Roos², Gilles S van Tienderen³, Kübra Köten³, Annelot Klaassen³, Wunan Mi³, Luc J W van der Laan³, Monique M A Verstegen⁴

Affiliations

¹ Department of Surgery, Erasmus MC Transplant Institute, University Medical Center Rotterdam, Rotterdam, the Netherlands. Electronic address: k.ober@erasmusmc.nl.
² Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, UK; Department of Surgery, University of Cambridge and NIHR Biomedical Research Centre, Cambridge, UK; Department of Surgery, Cambridge University Hospitals NHS Foundation Trust, Cambridge, UK.
³ Department of Surgery, Erasmus MC Transplant Institute, University Medical Center Rotterdam, Rotterdam, the Netherlands.
⁴ Department of Surgery, Erasmus MC Transplant Institute, University Medical Center Rotterdam, Rotterdam, the Netherlands. Electronic address: m.verstegen@erasmus.nl.

Abstract

Bile ducts are essential for bile transport and consist of complex branching tubular networks. Human patient-derived cholangiocyte develops a cystic rather than branching duct morphology. Here, we present a protocol to establish branching morphogenesis in cholangiocyte and cholangiocarcinoma organoids. We describe steps for the initiation, maintenance, and expansion of intrahepatic cholangiocyte organoids branching morphology. This protocol enables the study of organ-specific and mesenchymal-independent branching morphogenesis and provides an improved model to study biliary function and diseases. For complete details on the use and execution of this protocol, please refer to Roos et al. (2022).¹.

Keywords: Cancer; Cell Biology; Developmental biology; Organoids.

MeSH terms

Bile Duct Neoplasms* / pathology
Bile Ducts, Intrahepatic / pathology
Cholangiocarcinoma* / pathology
Humans
Morphogenesis
Organoids / pathology