Long-term expansion of human cells from liver and pancreas

Researchers of the University of Cambridge (UK) described, in the prestigious journal "Nature protocols", a method to grow human adult liver and pancreatic cells into self-assembling 3D structures that can sustain long-term expansion. 

In brief, after surgical extraction, liver tissues are minced into pieces and digested into a solution containing collagenase and dispase. After debris removal by filtration, cells are resuspended in a basement matrix (Matrigel) and overlaid in medium supplemented with EGF, FGF, HGF and Rspo1, which promotes Wnt signaling. In two weeks, 3D organoids should be visible and ready for passaging or cryopreservation. Cells of liver organoids can be subdivided, cryopreserved or differentiated into hepatocytes by removal of Rspo1 and addition of BMP7 and dexamethasone to culture medium. A similar protocol can be used to grow pancreatic cells by substitution of HGF with Noggin into culture medium.

The authors claim that 3D organoids can be very useful for studying liver and pancreas biology in a primary, non transformed, physiologically relevant system.

Link to journal page

3D Organoids of cells isolated from liver, on the left, or pancreas, on the right. Cells form 3D organoid structures after one week in culture. Images belong to figure 1 of the reference paper published on Nature Protocols.