Functional hepatobiliary organoids recapitulate liver development and reveal essential drivers of hepatobiliary cell fate determination

Abstract

During liver development, hepatocytes and cholangiocytes are concurrently differentiated from common liver progenitor cells and are assembled into hepatobiliary architecture to perform proper hepatic function. However, the generation of functional hepatobiliary architecture from hepatocytes in vitro is still challenging, and the exact molecular drivers of hepatobiliary cell lineage determination is largely unknown. In this study, functional hepatobiliary organoids (HBOs) are generated from hepatocytes. These HBOs contain a bile duct network surrounded by mature hepatocytes and stably maintain hepatic characteristics and function in vitro and upon transplantation in vivo. Morphological transition and expression profile of hepatocyte-derived organoids recapitulate the process of liver development. Gene regulation landscape of hepatocyte-derived organoids reveal that Tead4 and Ddit3 promote the cell fate commitment of liver progenitors to functional cholangiocytes and hepatocytes, respectively. Liver cell fate determination is reversed by inhibiting Tead4 or increasing Ddit3 expression both in vitro and upon transplantation in vivo. Collectively, hepatocyte-derived HBOs reveal the essential transcription drivers of liver hepatobiliary cell lineage determination and represent powerful models for liver development and regeneration.

Using Liver Organoids as Models to Study the Pathobiology of Rare Liver Diseases

“…Another approach is adding integrative cues to the co-culture system, and organoids can be controlled to provide a human-like microenvironment [71][72][73]. Engineering approaches to produce human-like organoid co-culture systems include adding mechanical cues, nutritional needs and metabolic cues, related tissues such as stromal and immune cells, and biosensing technology [71][72][73][74][75].…”

Cholangiocyte Organoids: The New Frontier in Regenerative Medicine for the Study and Treatment of Cholangiopathies

“…Advances in culture techniques and biomaterials hold promise for overcoming these challenges and unlocking the full potential of organoid research. A major limitation is the maintenance of cellular functionality and viability over time due to culture conditions, such as media composition and growth factors; as the volume of organoids increases, so does the demand for nutrients and oxygen [ 130 , 137 ]. At the same time, the accumulation of toxic metabolites causes cell death and necrosis.…”

Hepatocyte reprogramming in liver regeneration: Biological mechanisms and applications

“…A recent study employing random lineage tracing to quantify the clonal expansion of hepatocytes in a normal liver revealed that 84.1% of hepatocytes did not proliferate, and only a small number of hepatocytes generated new cells to maintain the mouse liver mass in 9.5-9.7 months [16]. However, once the liver is injured from partial hepatectomy or harmful stimuli such as APAP and DDC, the regenerative process is initiated by hepatocyte self-renewal or hepatocyte reprogramming [1,[27][28][29]. Due to the different stimulating factors of injury, hepatocytes choose different regeneration modes, taking the path to liver reconstruction or tumorgenesis (Fig.…”