Adult human hepatocyte organoid expansion for in vitro testing and in vivo engraftment

Abstract

The liver, an intricate organ with over 500 distinct functions within the human body, possesses remarkable regenerative capabilities following injury or resection. Its pivotal role in detoxification heightens its importance in determining the body's resilience to harmful substances. However, the rapid dedifferentiation of hepatocytes, the liver's primary parenchymal cells, in ex vivo conditions presents a challenge, hindering studies on liver function and regeneration in humans. To overcome this limitation, we have developed optimized in vitro models utilizing adult human hepatocyte organoids, which serve as valuable tools for modeling liver physiology and tissue engineering. Through these models, we investigated the effects of various growth factors on hepatic function, elucidated hepatocyte proliferation mechanisms, and systematically analyzed cell heterogeneity. Leveraging magnetic-activated cell sorting, we enhanced the functionality of hepatocyte organoids, prolonging their efficacy. We conducted in vivo implantation studies of human hepatocyte organoids, demonstrating their comparable functionality to freshly thawed hepatocytes. Notably, our organoids exhibited promising results in immunodeficient mouse models. Utilizing acute and chronic drug-induced liver injury models, our organoids suggest therapeutic potential in aiding patients' recovery from drug overdose. This innovative approach lays the groundwork for transitioning our hepatocyte organoid model toward clinical application, thereby advancing the prospect of utilizing these organoids as a therapeutic intervention for liver-related disorders.