Basaal II - P48 - Characterisation of peribiliary glands in a new model: precision-cut bile duct slices

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I.E.M. de Jong, A.P.M. Matton, R. Iswandana, S. Suriguga, T. van Haaften, J. Wiersema-Buist, D. Oosterhuis, T. Lisman, P. Olinga, R.J. Porte

Chair(s): prof. dr. Carla C. Baan, Erasmus MC Rotterdam

Thursday 10 march 2016

12:30 - 13:00h at Foyer

Categories: Postersessie

Parallel session: Postersessies XI - Opgesplitst in 3 tijdblokken en 3 categoriëen (klinisch, basaal, donatie)

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Non-anastomotic strictures (NAS) are a major cause of morbidity and graft failure after orthotopic liver transplantation. NAS is considered to be the most problematic biliary complication as it is the most frequent indication for retransplantation. Recent research suggests that the regenerative capacity of peribiliary glands (PBGs), which are niches of progenitor cells lining the bile ducts, plays a role in de development of NAS.  Precision-cut tissue slices (PCTS) are a widely used in vitro technique, in which thin slices of fresh human tissue are cut, incubated and cultured and thereby kept viable for up to several days. In PCTS, all the cell types of the tissue remain in their natural environment, with intercellular and cell-matrix interactions remaining intact. Our aim was to establish Precision-cut Bile Duct Slices (PCBDS) as a model to study the regenerative capacity of PBGs in human extrahepatic bile duct. 

Extra-hepatic bile ducts (EHBD) of livers declined for transplantation were used (n=5) and sliced using a Krumdieck tissue slicer. Thereafter, they were incubated in plates filled with Williams’ medium E (supplemented with glutamax, glucose and antibiotics; incubator with 80%O₂/20%CO₂ shaking at 90 rpm at 37°C). Medium was refreshed and slices were harvested every 24 hours for up to 144 hours. HE, PDX1, and CK19- stainings were performed on paraffin-embedded samples for histomorphological assessment. 

Histomorphological assessment showed cell loss gradually over time, conversely, PBGs and neural tissue survived for up to 144 hours. In addition, preliminary data show that these PBGs express PDX1, a progenitor marker, and CK19, a cholangiocyte marker. 

In conclusion, PCBDS allow for PBGs to remain viable up to 144 hours and express characteristic progenitor and cholangiocyte traits. These preliminary data suggest that PCBDS can be used to study PBGs and their regenerative capacity. Future research will focus on agents to stimulate regeneration of PBGs in this model, in the search of a potential intervention for NAS.